Patent application title: METHODS FOR DETERMINING RESPONSIVENESS TO MEK/ERK INHIBITORS
Inventors:
IPC8 Class: AC12Q16886FI
USPC Class:
1 1
Class name:
Publication date: 2020-04-23
Patent application number: 20200123622
Abstract:
A method for predicting the responsiveness of a cancer cell to an MEK
inhibitor, comprising detecting the presence of at least one mutation in
one or more genes selected from the group consisting of ADAM12, COL14A1,
TNN, and TP53, in the cancer cell, by contacting a nucleic acid sample
derived from the cancer cell with at least one oligonucleotide which
allows specific detection of the mutation; wherein presence of mutation
in ADAM12, COL14A1, TNN, TP53 and/or any combination thereof is
indicative of decreased responsiveness of the cancer cell to the ERK
inhibitor.Claims:
1) A method for treating a patient having cancer, comprising: obtaining a
sample derived from the patient; determining that the sample does not
have a mutation in TP53 gene which results in Q331R, C135fs, E285K,
V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice,
R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*,
E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in
TP53 protein; and administering an MEK inhibitor to the patient.
2) The method of claim 1, wherein the sample is a cancer cell or tissue derived from the patient.
3) The method of claim 1, wherein the MEK inhibitor is Trametinib.
4) The method of claim 1, wherein the determining step comprises amplifying at least a portion of the TP53 gene with an oligonucleotide as primer to generate an amplification product.
5) The method of claim 1, wherein the determining step comprises contacting the sample with an oligonucleotide which specifically hybridizes to the mutation of the TP53 gene to form a complex.
6) The method of claim 1, wherein the determining step involves an assay selected from the group consisting of sequencing, polymerase chain reaction (PCR), mass-spectrometric genotyping, HPLC, SSPC and a hybridization-based assay.
7) The method of claim 1, wherein the determining step further comprises determining if the patient has a second mutation in one or more genes selected from the group consisting of ADAM12, COL14A1 and TNN.
8) The method of claim 7, wherein the mutation in ADAM12 gene results in Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L or G308E in ADAM12 protein; wherein the mutation in COL14A1 gene results in R178W, L713 splice, Q1272K, L4791, L1295F, E1024K, P1467S, G737R, K1023T, G966C, S1512fs in COL14A1 protein; wherein the mutation in TNN gene results in V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN protein.
9) A method for treating a patient having cancer, comprising: obtaining a sample derived from the patient; determining that the sample has a mutation in TP53 gene which results in Q331R, C135fs, E285K, V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice, R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*, E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in TP53 protein; and administering a therapeutic agent to the patient, wherein the therapeutic agent is selected from the group consisting of a c-Met inhibitor, an agent targeting PI3K-Akt-mTOR signaling pathway, a chemotherapeutic agent, an anti-metabolite, an anti-hormonal agent, and an angiogenesis inhibitor.
10) The method of claim 9, wherein the sample is a cancer cell or tissue derived from the patient.
11) The method of claim 9, wherein the determining step comprises amplifying at least a portion of the TP53 gene with an oligonucleotide as primer to generate an amplification product.
12) The method of claim 9, wherein the determining step comprises contacting the sample with an oligonucleotide which specifically hybridizes to the mutation of the TP53 gene to form a complex.
13) The method of claim 9, wherein the determining step involves an assay selected from the group consisting of sequencing, polymerase chain reaction (PCR), mass-spectrometric genotyping, HPLC, SSPC and a hybridization-based assay.
14) The method of claim 9, wherein the determining step further comprises determining if the patient has a second mutation in one or more genes selected from the group consisting of ADAM12, COL14A1 and TNN.
15) The method of claim 14, wherein the mutation in ADAM12 gene results in Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L or G308E in ADAM12 protein; wherein the mutation in COL14A1 gene results in R178W, L713 splice, Q1272K, L4791, L1295F, E1024K, P1467S, G737R, K1023T, G966C, S1512fs in COL14A1 protein; wherein the mutation in TNN gene results in V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN protein.
Description:
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a divisional application of U.S. Ser. No. 15/301,379, which is the national phase stage of PCT/CN2015/075884 filed on Apr. 3, 2015, which claims the priority to Chinese patent application no. 201410135569.9, filed Apr. 4, 2014, the disclosure of which is incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
[0002] The present invention generally relates to methods for determining the responsiveness of a subject to treatment with a MEK or ERK inhibitor.
BACKGROUND OF THE INVENTION
[0003] The Ras-Raf-MEK-ERK signaling cascade (MEK/ERK pathway) is one of key pro-proliferation and pro-survival pathways. Mutations in the MEK/ERK pathway have been found to lead to uncontrolled growth in many cancers (e.g., melanoma). Compounds that inhibit steps in the MEP/ERK pathway have been used to treat cancer. However, some patients that harbor mutations in MEK/ERK pathway show resistance to MEK or ERK inhibitors.
[0004] There is a need for an effective means of determining which patients having mutations in MEK/ERK pathway will resist to treatment of MEK or ERK inhibitors and for incorporating such determination into effective treatment.
BRIEF SUMMARY OF THE INVENTION
[0005] In one aspect, the present disclosure provides a method for predicting the responsiveness of a cancer cell to an MEK inhibitor. In certain embodiments, the method comprises detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, and TP53, in the cancer cell, by contacting a nucleic acid sample derived from the cancer cell with at least one oligonucleotide which allows specific detection of the mutation; wherein presence of mutation in ADAM12, COL14A1, TNN, TP53 and/or any combination thereof is indicative of decreased responsiveness of the cancer cell to the MEK inhibitor.
[0006] In certain embodiments, the cancer cell is derived from a cancer patient.
[0007] In certain embodiments, the MEK inhibitor is Trametinib.
[0008] In certain embodiments, the mutation in ADAM12 is selected from the group consisting of mutation Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L and G308E in ADAM12; the mutation in COL14A1 is selected from the group consisting of mutation R178W, L713 splice, Q1272K, L479I, L1295F, E1024K, P1467S, G737R, K1023T, G966C, S1512fs in COL14A1; the mutation in TNN is selected from the group consisting of mutation V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN; the mutation in TP53 is selected from the group consisting of mutation Q331R, C135fs, E285K, V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice, R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*, E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in TP53.
[0009] In certain embodiments, the detecting step comprises amplifying at least a portion of the gene with the oligonucleotide as primer, and detecting the amplification product and thereby determining the presence of the mutation in the gene.
[0010] In certain embodiments, the detecting step comprises contacting the nucleic acid sample with the oligonucleotide which specifically hybridizes to the mutation of the gene to form a complex, and detecting the formation of the complex and thereby determining the presence of the mutation in the gene.
[0011] In anther aspect, the present disclosure provides a method of identifying a likely responder or a likely non-responder to an MEK inhibitor, comprising detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, and TP53, in a sample from the patient, by contacting the sample with at least one oligonucleotide which allows specific detection of the mutation; identifying the patient as a likely non-responder to the MEK inhibitor if at least one mutation in in ADAM12, COL14A1, TNN, TP53 and/or any combination thereof is detected in the sample.
[0012] In certain embodiments, the MEK inhibitor is Trametinib.
[0013] In certain embodiments, the mutation in ADAM12 is selected from the group consisting of mutation Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L and G308E in ADAM12; the mutation in COL14A1 is selected from the group consisting of mutation R178W, L713 splice, Q1272K, L479I, L1295F, E1024K, P1467S, G737R, K1023T, G966C, S1512fs in COL14A1; the mutation in TNN is selected from the group consisting of mutation V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN; the mutation in TP53 is selected from the group consisting of mutation Q331R, C135fs, E285K, V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice, R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*, E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in TP53.
[0014] In certain embodiments, the method further comprises recommending the patient who is identified as a likely non-responder not to be treated with a monotherapy of the ERK inhibitor, or not to be treated with an MEK inhibitor.
[0015] In certain embodiments, the method further comprises recommending the patient who is identified as a likely non-responder to be treated with a different MEK inhibitor, or to be treated with a combined therapy of a different MEK inhibitor and an additional therapeutic agent of distinct mechanism.
[0016] In certain embodiments, the method further comprises recommending the patient who is identified as a likely responder to be treated with the MEK inhibitor.
[0017] In certain embodiments, the sample is a cancer cell or tissue derived from the patient.
[0018] In another aspect, the present disclosure provides a method for predicting the responsiveness of a cancer cell to a ERK inhibitor, comprising detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, PEX5L, TNN and TP53, in the cancer cell, by contacting a nucleic acid sample derived from the cancer cell with at least one oligonucleotide which allows specific detection of the mutation; wherein presence of the mutation in ADAM12, PEX5L, TNN, TP53 and/or any combination thereof is indicative of decreased responsiveness of the cancer cell to the ERK inhibitor.
[0019] In certain embodiments, the cancer cell is derived from a cancer patient.
[0020] In certain embodiments, the ERK inhibitor is SCH772984.
[0021] In certain embodiments, the mutation in ADAM12 is selected from the group consisting of mutation Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L and G308E in ADAM12; the mutation in PEX5L is selected from the group consisting of mutation D179N, S229Y, G4E, T89K, Q355E, D39N, L571F, D113N in PEX5L; the mutation in TNN is selected from the group consisting of mutation V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN; the mutation in TP53 is selected from the group consisting of mutation Q331R, C135fs, E285K, V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice, R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*, E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in TP53.
[0022] In certain embodiments, the detecting step comprises amplifying at least a portion of the gene with the oligonucleotide as primer, and detecting the amplification product and thereby determining the presence of the mutation in the gene.
[0023] In certain embodiments, the detecting step comprises contacting the nucleic acid sample with the oligonucleotide which specifically hybridizes to the mutation of the gene to form a complex, and detecting the formation of the complex and thereby determining the presence of the mutation in the gene.
[0024] In yet another aspect, the present disclosure provides a method of identifying a likely responder or a likely non-responder to an ERK inhibitor, comprising detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, PEX5L, TNN and TP53, in a sample from the patient, by contacting the sample with at least one oligonucleotide which allows specific detection of the mutation; identifying the patient as a likely non-responder to the ERK inhibitor if at least one mutation in ADAM12, PEX5L, TNN, TP53 and/or any combination thereof is detected in the sample.
[0025] In certain embodiments, the ERK inhibitor is SCH772984.
[0026] In certain embodiments, the mutation in ADAM12 is selected from the group consisting of mutation Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L and G308E in ADAM12; the mutation in PEX5L is selected from the group consisting of mutation D179N, S229Y, G4E, T89K, Q355E, D39N, L571F, D113N in PEX5L; the mutation in TNN is selected from the group consisting of mutation V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN; the mutation in TP53 is selected from the group consisting of mutation Q331R, C135fs, E285K, V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice, R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*, E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in TP53.
[0027] In certain embodiments, the method further comprises recommending the patient who is identified as a likely non-responder not to be treated with a monotherapy of the ERK inhibitor, or not to be treated with an ERK inhibitor.
[0028] In certain embodiments, the method further comprises recommending the patient who is identified as a likely non-responder to be treated with a different ERK inhibitor, or to be treated with a combined therapy of a different ERK inhibitor and an additional therapeutic agent of distinct mechanism.
[0029] In certain embodiments, the sample is a cancer cell or tissue derived from the patient.
[0030] In another aspect, the present disclosure provides a kit comprising at least one oligonucleotide useful for determining the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, TP53, ITGB, and PEX5L.
[0031] In certain embodiments, the at least one oligonucleotide comprises a first oligonucleotide useful for determining the presence of at least one mutation in ADAM12, a second oligonucleotide useful for determining the presence of at least one mutation in COL14L1, a third oligonucleotide useful for determining the presence of at least one mutation in TNN, a fourth oligonucleotide useful for determining the presence of at least one mutation in TP53, or any combination thereof.
[0032] In certain embodiments, the at least one oligonucleotide comprises a first oligonucleotide useful for determining the presence of at least one mutation in ADAM12, a second oligonucleotide useful for determining the presence of at least one mutation in PEX5L, a third oligonucleotide useful for determining the presence of at least one mutation in TNN, a fourth oligonucleotide useful for determining the presence of at least one mutation in TP53, and or combination thereof.
[0033] In certain embodiments, the at least one oligonucleotide comprises a pair of primer useful for amplifying at least a portion of the gene sequence, or comprises a probe useful for specifically hybridizing to the mutation of the gene to form a complex.
[0034] In another aspect, the present disclosure provides use of at least one oligonucleotide in the manufacture of a kit for predicting the responsiveness of a cancer cell or a cancer patient to an MEK inhibitor or a ERK inhibitor, wherein the oligonucleotide is useful for detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, TP53, and PEX5L.
BRIEF DESCRIPTION OF THE FIGURES
[0035] FIG. 1 illustrates the increased sensitivity to MEK inhibitor Trametinib in cells harboring mutations in ADAM12 gene.
[0036] FIG. 2 illustrates the decreased sensitivity to MEK inhibitor Trametinib in cells harboring mutations in COL14A1 gene.
[0037] FIG. 3 illustrates the increased sensitivity to MEK inhibitor Trametinib in cells harboring mutations in TNN gene.
[0038] FIG. 4 illustrates the increased sensitivity to MEK inhibitor Trametinib in cells harboring mutations in TP53 gene.
[0039] FIG. 5 illustrates the increased sensitivity to MEK inhibitor Trametinib in cells harboring multiple mutations in ADAM12, COL14A1, TNN, and TP53 gene.
[0040] FIG. 6 illustrates the increased sensitivity to ERK inhibitor SCH772984 in cells harboring mutations in ADAM12 gene.
[0041] FIG. 7 illustrates the increased sensitivity to ERK inhibitor SCH772984 in cells harboring mutations in PEX5L gene.
[0042] FIG. 8 illustrates the increased sensitivity to ERK inhibitor SCH772984 in cells harboring mutations in TNN gene.
[0043] FIG. 9 illustrates the increased sensitivity to ERK inhibitor SCH772984 in cells harboring mutations in TP53 gene.
[0044] FIG. 10 illustrates the increased sensitivity to ERK inhibitor SCH772984 in cells harboring multiple mutations in ADAM 12, PEX5L, TNN and TP53 gene.
DETAILED DESCRIPTION OF THE INVENTION
[0045] In one aspect, the present disclosure provides a method for predicting the responsiveness of a cancer cell to an MEK inhibitor. In certain embodiments, the method comprises detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, and TP53, in the cancer cell, by contacting a nucleic acid sample derived from the cancer cell with at least one oligonucleotide which allows specific detection of the mutation; wherein presence of mutation in ADAM12, COL14A1, TNN, TP53 and/or any combination thereof is indicative of decreased responsiveness of the cancer cell to the ERK inhibitor.
[0046] The mitogen-activated extracellular signal-regulated kinase (MEK)-extracellular regulated protein kinases (ERK) cascade, also known as Ras-Raf-MEK-ERK signaling pathway, is one of the key signaling pathways involved in tumor oncogenic growth and progression. The signal pathway starts when a signaling molecule (e.g., a growth factor) binds to the receptor on the cell surface. This triggers Ras (a GTPase) to sap its GDP for a GTP. The GTP-bound Ras then activate Raf, which activates MEK, which activates ERK. ERK then activates some proteins, such as myc, that control cell division and cell survival. When one or more proteins in the pathway, such as Ras, are mutated, it can lead to the signaling pathway stuck in the activated status, which is a necessary step in the development of many cancers. As a result, inhibitors of the MEK/ERK signaling pathway have been developed to treat cancer. Certain patients have been found resistant to MEK or ERK inhibitors. The mechanisms underlying resistance to these inhibitors are unclear.
[0047] Multiple MEK and ERK1/2 inhibitors are currently under clinical investigation for cancer treatment and more agents targeting MEK or ERK1/2 are under preclinical development. Examples of MEK inhibitors include without limitation Trametinib (GSK1120212), Selumetinib, Binimetinib (MEK162), PD-325901, Cobimetinib (GDC-0973, XL518), and CI-1040 (PD035901). ERK inhibitors include without limitation SCH772984, FR180204, GDC-0994.
[0048] In certain embodiments, the MEK inhibitor is Trametinib. Trametinib (trade name Mekinist) has chemical name N-(3-{3-Cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-t- rioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H)-yl}phenyl)acetamide. The structure of Trametinib is illustrated below.
##STR00001##
[0049] As used herein, the term "responsiveness" refers to the likeliness of a cell/individual/patient/subject responding to the treatment of MEK or ERK inhibitor, i.e. showing decreased proliferation/growth and/or increased cell death after being treated with a MEK or ERK inhibitor. In certain embodiments, the responsiveness can be scaled as insensitive (i.e., less likely to respond), sensitive (likely to respond) and uncertain. In certain embodiments, the cell/individual/patient/subject is less likely to respond to a treatment of MEK or ER inhibitor when the cell/individual/patient/subject shows a decreased likeliness that a pathological complete response (pcR), i.e. absence of invasive cancer, will occur. In certain embodiments, the deceased likeliness means about 70%, 60%, 50%, 40%, 30%, 20%, 10% likeliness of the pcR occurred in a reference patient (e.g., a patient without mutation in the gene of interest). In certain embodiments, responsiveness of a cell can be evaluated by measuring IC50 of the cell to a MEK or ERK inhibitor.
[0050] As used herein, the term "mutation" refers to the deviation of a genomic DNA from a normal reference (e.g., wild-type genomic DNA), for example, additions, deletions, insertions, rearrangements, inversions, transitions, transversions, frame-shift mutations, nonsense mutations, missense mutations, translocations, and single nucleotide polymorphisms.
[0051] Methods of detecting the presence of a mutation in a gene are described herein and known in the art (in general, see e.g., Molecular Cloning A Laboratory Manual, 2nd Ed., ed. By Sambrook, Fritsch and Maniatis, Cold Spring Harbor Laboratory Press, 1989). Examples of the method include, without limitation, sequencing of nucleic acids (e.g., Sanger di-deoxy sequencing, "next generation" sequencing methods and single molecule sequencing), PCR (polymerase chain reaction)-based assay (real-time RCR, PCR-RFLP assay (see Cancer Research 59 (1999), 5169-5175), mass-spectrometric genotyping (e.g. MALDI-TOF), HPLC, enzymatic methods and SSPC (single strand conformation polymorphism analysis (see Pathol Int (1996) 46, 801-804)), hybridization-based assay (e.g., Northern-blot, Southern blot, 5'-exonuclease (TaqMan.TM.) probe, molecular beacons, fluorescence energy transfer probes, Scorpion probes).
[0052] In certain embodiments, the method may include enzymatic amplification of DNA or cDNA fragments of the gene to be evaluated by PCR. The resulting PCR products may be subjected to either conventional Sanger-based dideoxy nucleotide sequencing methods or parallel sequencing methods ("next generation sequencing") such as those marketed by Roche (454 technology), Illumina (Solexa technology) ABI (Solid technology) or Invitrogen (IonTorrent). Mutations may be identified from sequence reads by comparison with publicly available gene sequence databases. Alternatively, mutations may be identified by incorporation of allele-specific probes that can either be detected using enzymatic detection reactions, fluorescence, mass spectrometry or others.
[0053] In certain embodiments, the method may include amplifying DNA or cDNA fragments of the gene of interest using primers specifically bind to only one of normal and mutated sequence. As a result, amplification product can only be found in one of normal and mutated genes. As such, the presence of the mutation in the gene can be determined by detecting the presence of the amplification product.
[0054] In certain embodiments, the method may include contacting the nucleic acid sample with an oligonucleotide probe which specifically hybridizes to the mutation of the gene to form a complex. The oligonucleotide probe can be designed as not hybridizing to the normal sequence of the gene. The presence of the hybridization complex can be detected using reporter signals, e.g., fluorescence. As a result, the presence of the mutation in the gene can be determined by detecting the formation of the complex.
[0055] In certain embodiments, the mutation is present in an exon of the gene. In certain embodiment, the presence of the mutation leads to the amino acid change of the polypeptide encoded by the gene. Table 3 shows exemplary nucleic acid sequences of the mutations to be determined in accordance to the present invention. As used herein, a specific mutation is annotated as the resulted amino acid change. For example, mutation Q650K refers to a codon/triplet encoding amino acid K at position 650 of the gene, where amino acid G exists in wild type sequence.
[0056] ADAM12 gene (Gene ID: 8038) encodes a member of the ADAM (a disintegrin and metaloprotease) protein family. Members of the ADAM family are membrane-anchored proteins structurally related to snake venom disintegrins, and have been found involved in cell-cell and cell-matrix interactions. ADAM12 gene has two alternative spliced transcripts: a shorter secreted form and a longer member-bound form.
[0057] COL14A1 gene (Gene ID: 7373) encodes the alpha chain of type XIV collage, a member of the FACIT (fibril-associated collagens with interrupted triple helices) collagen family. Type XIV collagen interacts with the fibril surface and is involved in the regulation of fribrillogenesis.
[0058] TNN gene (Gene ID: 63923) encodes tenascin N precursor. Tenascin is a family of extracellular matrix glycoproteins, whose member has been found to in healing wounds and in the stroma of some tumors.
[0059] TP53 gene (Gene ID: 7157) encodes tumor protein p53, which is a tumor suppressor protein containing transcriptional activation, DNA binding, and oligomerization domains. Tumor protein p53 responds to diverse cellular stresses to regulate expression of target genes, thereby inducing cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. Mutations in TP 53 gene have been found to associate with a variety of cancers. Alternative splicing of TP53 gene and the use of alternate promoters result in multiple transcript variants and isoforms. Additional isoforms have also been shown to result from the use of alternate translation initiation codons. As used herein, position number refers to the sequence of tumor protein p53 isoform a.
[0060] In certain embodiments, the mutation in ADAM12 is selected from the group consisting of mutation Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L and G308E in ADAM12; the mutation in COL14A1 is selected from the group consisting of mutation R178W, L713 splice, Q1272K, L479I, L1295F, E1024K, P1467S, G737R, K1023T, G966C, S1512fs in COL14A1; the mutation in TNN is selected from the group consisting of mutation V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN; the mutation in TP53 is selected from the group consisting of mutation Q331R, C135fs, E285K, V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice, R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*, E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in TP53, wherein "*" means the mutation leads to a stop condon, "fs" means the mutations leads to frame shift, "splice" means the mutation leads to alternative splice of mRNA.
[0061] The cancer cell maybe, for example, derived from lung cancer, non small cell lung (NSCL) cancer, bronchioloalviolar cell lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, gastric cancer, colon cancer, breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, mesothelioma, hepatocellular cancer, biliary cancer, chronic or acute leukemia, lymphocytic lymphomas, neoplasms of the central nervous system (CNS), spinal axis tumors, brain stem glioma, glioblastoma multiforme, astrocytomas, schwanomas, ependymonas, medulloblastomas, meningiomas, squamous cell carcinomas, pituitary adenoma, including refractory versions of any of the above cancers, or a combination of one or more of the above cancers. In certain embodiments, the cancer cell is derived from a cancer patient.
[0062] In another aspect, the present disclosure provides a method of identifying a likely responder or a likely non-responder to an MEK inhibitor, comprising detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, and TP53, in a sample from the patient, by contacting the sample with at least one oligonucleotide which allows specific detection of the mutation; identifying the patient as a likely non-responder to the MEK inhibitor if at least one mutation in ADAM12, COL14A1, TNN, TP53 and/or any combination thereof is detected in the sample.
[0063] As used herein, the term "responder" can refer to an individual/patient/subject that is more likely to respond to a treatment using a MEK or ERK inhibitor. "More likely to respond" as used herein refers to an increased likeliness that a pathological complete response will occur in a patient treated with a MEK or ERK inhibitor. The term "non-responder" can refer to an individual/patient/subject that is less likely to respond to a treatment using a MEK or ERK inhibitor. "Less likely to respond" as used herein refers to an decreased likeliness that a pathological complete response will occur in a patient treated with a MEK or ERK inhibitor.
[0064] In certain embodiments, in cases where it is assessed that the patient is a likely "responder," said patient is recommended to be treated with an MEK or ERK inhibitor.
[0065] In cases where the patient is identified as a likely non-responder, said patient is recommended not to be treated with a monotherapy of the MEK or ERK inhibitor, or not to be treated with an MEK or ERK inhibitor.
[0066] In certain embodiments, wherein the patient is identified as a likely non-responder to a MEK inhibitor, the patient is recommended to be treated with a different MEK inhibitor, or to be treated with a combined therapy of a different MEK inhibitor and an additional therapeutic agent of distinct mechanism. Examples of an MEK inhibitor different from Trametinib include, without limitation, Selumetinib, Binimetinib (MEK162), PD-325901, Cobimetinib (GDC-0973, XL518), and CI-1040 (PD035901).
[0067] In certain embodiments, the additional therapeutic agent of distinct mechanism can be an agent targeting PI3K-Akt-mTOR signaling pathway. The agents targeting PI3K-Akt-mTOR signaling pathway are known in the art and comprise, without limitation, fused pyrimidine derivatives as disclosed in U.S. Pat. No. 8,022,205 B2 or fused pyrrolopyrimidine derivatives as disclosed in WO2009/099163.
[0068] In certain embodiments, the additional agent of distinct mechanism can include c-Met inhibitors (e.g., ARQ197 (taventinib, developed by Daichi Sankyo and ArQule), AMG458 (developed by Amgen), GSK1363089 (also known as XL880 or foretinib, developed GSK), crizotinib (also known as PF2341066, developed by Pfizer), PF04217903 (developed by Pfizer), INCB28060 (developed by Incyte), E7050 (developed by Eisai), MK-2461 (developed by Merck), BMS-777607 (developed by BMS), JNJ-38877605 (developed by Johnson & Johnson), XL184 (developed by BMS/Exelixis)).
[0069] In certain embodiments, the additional therapeutic agent of distinct mechanism can be chemotherapeutic agents (e.g., cyclophosphamide (CTX; e.g. Cytoxan.RTM.), chlorambucil (CHL; e.g. Leukeran.RTM.), cisplatin (CisP; e.g. Platinol.RTM.) busulfan (e.g. Myleran.RTM.), melphalan, carmustine (BCNU), streptozotocin, triethylenemelamine (TEM), mitomycin C)
[0070] In certain embodiments, the additional therapeutic agent of distinct mechanism can be anti-metabolites, such as methotrexate (MTX), etoposide (VP16; e.g. Vepesid.RTM.), 6-mercaptopurine (6MP), 6-thiocguanine (6TG), cytarabine (Ara-C), 5-fluorouracil (5-FU), capecitabine (e.g. Xeloda.RTM.), dacarbazine (DTIC)).
[0071] In certain embodiments, the additional therapeutic agent of distinct mechanism can be other antitumor agents, such as paclitaxel (e.g. Taxol.RTM.) and pactitaxel derivatives, the cytostatic agents, glucocorticoids such as dexamethasone (DEX; e.g. Decadron.RTM.) and corticosteroids such as prednisone, nucleoside enzyme inhibitors such as hydroxyurea, amino acid depleting enzymes such as asparaginase, leucovorin, folinic acid, raltitrexed, and other folic acid derivatives, and similar, diverse antitumor agents.
[0072] In certain embodiments, the additional therapeutic agent of distinct mechanism can be anti-hormonal agents (e.g., steroid receptor antagonists, anti-estrogens such as tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, other aromatase inhibitors, 42-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, and toremifene (e.g. Fareston.RTM.); anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above; agonists and/or antagonists of glycoprotein hormones such as follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), and luteinizing hormone (LH) and LHRH (leuteinizing hormone-releasing hormone); the LHRH agonist goserelin acetate, commercially available as Zoladex.RTM. (AstraZeneca); the LHRH antagonist D-alaninamide N-acetyl-3-(2-naphthalenyl)-D-alanyl-4-chloro-D-phenylalanyl-3-(3-pyridin- yl)-D-alanyl-L-seryl-N6-(3-pyridinylcarbonyl)-L-lysyl-N6-(3-pyridinylcarbo- nyl)-D-lysyl-L-leucyl-N6-(1-methylethyl)-L-lysyl-L-proline (e.g Antide.RTM., Ares-Serono); the LHRH antagonist ganirelix acetate; the steroidal anti-androgens cyproterone acetate (CPA) and megestrol acetate, commercially available as Megace.RTM. (Bristol-Myers Oncology); the nonsteroidal anti-androgen flutamide (2-methyl-N-[4, 20-nitro-3-(trifluoromethyl)phenylpropanamide), commercially available as Eulexin.RTM. (Schering Corp.); the non-steroidal anti-androgen nilutamide, (5,5-dimethyl-3-[4-nitro-3-(trifluoromethyl-4'-nitrophenyl)-4,4-dimethyl-- imidazolidine-dione); and antagonists for other non-permissive receptors, such as antagonists for RAR, RXR, TR, VDR, and the like).
[0073] In certain embodiments, the additional therapeutic agent of distinct mechanism can be angiogenesis inhibitors (e.g., VEGFR inhibitors, such as SU-5416 and SU-6668 (Sugen Inc. of South San Francisco, Calif., USA), or as described in, for example International Application Nos. WO 99/24440, WO 99/62890, WO 95/21613, WO 99/61422, WO 98/50356, WO 99/10349, WO 97/32856, WO 97/22596, WO 98/54093, WO 98/02438, WO 99/16755, and WO 98/02437, and U.S. Pat. Nos. 5,883,113, 5,886,020, 5,792,783, 5,834,504 and 6,235,764; VEGF inhibitors such as IM862 (Cytran Inc. of Kirkland, Wash., USA); angiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colo.) and Chiron (Emeryville, Calif.); and antibodies to VEGF, such as bevacizumab (e.g. Avastin.TM. Genentech, South San Francisco, Calif.), a recombinant humanized antibody to VEGF; integrin receptor antagonists and integrin antagonists, such as to .alpha..sub.v.beta..sub.3, .alpha..sub.v.beta..sub.35 and .alpha..sub.v.beta..sub.6 integrins, and subtypes thereof, e.g. cilengitide (EMD 121974), or the anti-integrin antibodies, such as for example .alpha..sub.v.beta..sub.33 specific humanized antibodies (e.g. Vitaxin.RTM.); factors such as IFN-alpha (U.S. Pat. Nos. 41,530,901, 4,503,035, and 5,231,176); angiostatin and plasminogen fragments (e.g. kringle 14, kringle 5, kringle 1-3 (O'Reilly, M. S. et al. (1994) Cell 79:315-328; Cao et al. (1996) J. Biol. Chem. 271: 29461-29467; Cao et al. (1997) J. Biol. Chem. 272:22924-22928); endostatin (O'Reilly, M. S. et al. (1997) Cell 88:277; and International Patent Publication No. WO 97/15666); thrombospondin (TSP-1; Frazier, (1991) Curr. Opin. Cell Biol. 3:792); platelet factor 4 (PF4); plasminogen activator/urokinase inhibitors; urokinase receptor antagonists; heparinases; fumagillin analogs such as TNP-4701; suramin and suramin analogs; angiostatic steroids; bFGF antagonists; flk-1 and flt-1 antagonists; anti-angiogenesis agents such as MMP-2 (matrix-metalloprotienase 2) inhibitors and MMP-9 (matrix-metalloprotienase 9) inhibitors).
[0074] In certain embodiments, the sample is a cancer cell or tissue derived from the patient.
[0075] In another aspect, the present disclosure provides a method for predicting the responsiveness of a cancer cell to a ERK inhibitor, comprising detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, PEX5L, TNN and TP53, in the cancer cell, by contacting a nucleic acid sample derived from the cancer cell with at least one oligonucleotide which allows specific detection of the mutation; wherein presence of the mutation in ADAM12, PEX5L, TNN, TP53 and/or any combination thereof is indicative of decreased responsiveness of the cancer cell to the ERK inhibitor.
[0076] In certain embodiments, the cancer cell is derived from a cancer patient.
[0077] In certain embodiments, the ERK inhibitor is SCH772984. SCH772984, with chemical name (R)-1-(2-oxo-2-(4-(4-(pyrimidin-2-yl)phenyl)piperazin-1-yl)ethyl)-N-(3-(p- yridin-4-yl)-1H-indazol-5-yl)pyrrolidine-3-carboxamide, is a novel, selective and ATP competitive inhibitor of ERK1/2 (see Morris E J et al., Discovery of a novel ERK inhibitor with activity in models of acquired resistance to BRAF and MEK inhibitors, Cancer Discov. 20133(7): 742-50). The structure of SCH772984 is illustrated as below.
##STR00002##
[0078] In certain embodiments, the mutation in ADAM12 is selected from the group consisting of mutation Q650K, R240L, C440Y, Q228E, H247D, M322I, T97fs, P168L and G308E in ADAM12; the mutation in PEX5L is selected from the group consisting of mutation D179N, S229Y, G4E, T89K, Q355E, D39N, L571F, D113N in PEX5L; the mutation in TNN is selected from the group consisting of mutation V353M, Y296S, A733P, D707Y, D471Y, P1010T, S71L, D457Y, P1155L, R476C, Q872H, Q261L, D798Y, C1237*, D67N and T823S in TNN; the mutation in TP53 is selected from the group consisting of mutation Q331R, C135fs, E285K, V274F, Y220C, P250L, R175H, R248Q, R280K, R248L, C176Y, A307_splice, R273L, R158L, A138fs, H193R, A159D, C277F, R248W, Y220C, V274F, R196*, E224_splice, K164*, M246I, A159V, S241F, C242R, S261_splice, E339* in TP53.
[0079] In certain embodiments, the detecting step comprises amplifying at least a portion of the gene with the oligonucleotide as primer, and detecting the amplification product and thereby determining the presence of the mutation in the gene.
[0080] In certain embodiments, the detecting step comprises contacting the nucleic acid sample with the oligonucleotide which specifically hybridizes to the mutation of the gene to form a complex, and detecting the formation of the complex and thereby determining the presence of the mutation in the gene.
[0081] In yet another aspect, the present disclosure provides a method of identifying a likely responder or a likely non-responder to an ERK inhibitor, comprising detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, PEX5L, TNN and TP53, in a sample from the patient, by contacting the sample with at least one oligonucleotide which allows specific detection of the mutation; identifying the patient as a likely non-responder to the ERK inhibitor if at least one mutation in ADAM12, PEX5L, TNN, TP53 and/or any combination thereof is detected in the sample.
[0082] In certain embodiments, the method further comprises recommending the patient who is identified as a likely non-responder not to be treated with a monotherapy of the ERK inhibitor, or not to be treated with an ERK inhibitor.
[0083] In certain embodiments, the method further comprises recommending the patient who is identified as a likely non-responder to be treated with a different ERK inhibitor, or to be treated with a combined therapy of a different ERK inhibitor and an additional therapeutic agent of distinct mechanism. Examples of ERK inhibitors other than SCH772984 include FR180204, GDC-0994.
[0084] In another aspect, the present disclosure provides a kit comprising at least one oligonucleotide useful for determining the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, TP53, ITGB, and PEX5L.
[0085] In certain embodiments, the at least one oligonucleotide comprises a first oligonucleotide useful for determining the presence of at least one mutation in ADAM12, a second oligonucleotide useful for determining the presence of at least one mutation in COL14L1, a third oligonucleotide useful for determining the presence of at least one mutation in TNN, a fourth oligonucleotide useful for determining the presence of at least one mutation in TP53, or any combination thereof.
[0086] In certain embodiments, the at least one oligonucleotide comprises a first oligonucleotide useful for determining the presence of at least one mutation in ADAM12, a second oligonucleotide useful for determining the presence of at least one mutation in PEX5L, a third oligonucleotide useful for determining the presence of at least one mutation in TNN, a fourth oligonucleotide useful for determining the presence of at least one mutation in TP53, and or combination thereof.
[0087] In certain embodiments, the at least one oligonucleotide comprises a pair of primer useful for amplifying at least a portion of the gene sequence, or comprises a probe useful for specifically hybridizing to the mutation of the gene to form a complex.
[0088] In another aspect, the present disclosure provides use of at least one oligonucleotide in the manufacture of a kit for predicting the responsiveness of a cancer cell or a cancer patient to an ERK inhibitor or a MEK inhibitor, wherein the oligonucleotide is useful for detecting the presence of at least one mutation in one or more genes selected from the group consisting of ADAM12, COL14A1, TNN, TP53, and PEX5L.
Example 1
[0089] The following is an example of identifying genes correlated with sensitivity to MEK inhibitors and/or ERK inhibitors.
[0090] We examined the anti-proliferation activity of a MEK inhibitor, trametinib, and an ERK1/2 inhibitor, SCH772984, in a panel of 50 cell lines (see Table 1).
TABLE-US-00001 TABLE 1 Cell lines used in the screen Cancer Ras/raf Type No. Cell line mutation Growth P. Medium 1. 1.1 BT474 150 Adherent DMEM + 0.01 mg/ml bovine insulin Breast 1.2 DU4475 423 Suspension RPMI-1640 1.3 MDA-MB-231 576 Adherent L15 1.4 ZR-75-1 493 Adherent RPMI-1640 2. 2.1 COLO 205 591 Adherent RPMI-1640 Colorectal 2.2 DLD-1 164 Adherent RPMI-1640 2.3 HCT-116 444 Adherent McCoys'5a 2.4 HCT-15 627 Adherent RPMI-1640 2.5 HCT-8 354 Adherent RPMI-1640 2.6 HT-29 504 Adherent McCoys'5a 2.7 KM12 L4 779 Adherent DMEM 2.8 LoVo 737 Adherent F12K 2.9 LS513 775 Adherent RPMI-1640 2.1 RKO 44 Adherent MEM 2.11 SW1116 42 Adherent L15 2.12 SW480 237 Adherent L15 2.13 SW620 590 Adherent L15 3. 3.1 Hep G2 243 Adherent EMEM Liver 3.2 HuCCT1 396 Adherent RPMI-1640 3.3 SK-HEP-1 171 Adherent MEM + 0.1 mMNEAA 3.4 SNU-387 287 Adherent RPMI-1640 4. 4.1 A549 101 Adherent F12K Lung 4.2 Calu-6 254 Adherent EMEM 4.3 NCI-H1155 584 Suspension ACL-4 4.4 NCI-H1299 577 Adherent RPMI-1640 4.5 NCI-H1373 360 Adherent RPMI-1640 4.6 NCI-H1395 386 Adherent RPMI-1640 4.7 NCI-H1573 426 Adherent ACL-4 4.8 NCI-H1651 429 Adherent ACL-4 4.9 NCI-H1666 425 Adherent ACL-4 4.1 NCI-H1792 400 Adherent RPMI-1640 4.11 NCI-H2009 513 Adherent HITES + 10% FBS 4.12 NCI-H2227 501 Adherent HITES + 10% FBS 4.13 NCI-H23 380 Adherent RPMI-1640 4.14 NCI-H358 571 Adherent RPMI-1640 4.15 NCI-H441 759 Adherent RPMI-1640 4.16 NCI-H460 24 Adherent RPMI-1640 4.17 SK-LU-1 403 Adherent EMEM 4.18 SW1271 471 Adherent L15 5. 5.1 AsPC-1 736 Adherent RPMI-1640 Pancreas 5.2 Capan-1 731 Adherent IMDM + 20% FBS 5.3 CFPAC-1 43 Adherent IMDM + 20% FBS 5.4 MIA PaCa-2 167 Adherent DMEM + 10% FBS + 2.5% HS 5.5 PANC-1 156 Adherent DMEM 6. 6.1 PL45 422 Adherent DMEM Skin 6.2 A2058 420 Adherent DMEM 6.3 A-375 716 Adherent DMEM 6.4 SK-MEL-5 154 Adherent MEM + 10% FBS + 0.01 mM NEAA 7. 7.1 AGS 295 Adherent F12K Stomach 7.2 SNU-1 292 Suspension RPMI-1640 7.3 SNU-719 552 Adherent RPMI-1640
[0091] Materials and Methods
[0092] Cell Culture
[0093] All the cells will be cultured in the media supplemented with 10% FBS except for which are marked specially, in the temperature of 37.degree. C., 5% CO.sub.2 and 95% humidity.
[0094] Cell Viability Reagent
[0095] Cell viability is assayed by using CellTiter-Glo.RTM. Luminescent Cell Viability Assay Kit (Cat. No.: G7572, Promega. Store at -20.degree. C.). To prepare the CellTiter_Glo Reagent, the CellTiter-Glo Buffer was thawed and equilibrated to room temperature prior to use. For convenience the CellTiter-Glo Buffer may be thawed and stored at room temperature for up to 48 hours prior to use. The lyophilized CellTiter-Glo Substrate is equilibrated to room temperature prior to use. The appropriate volume (100 ml) of CellTiter-Glo Buffer is transferred into the amber bottle containing CellTiter-Glo Substrate to reconstitute the lyophilized enzyme/substrate mixture, which forms the CellTiter-Glo Reagent. In certain cases, the entire liquid volume of the CellTiter-Glo Buffer bottle may be added to the CellTiter-Glo Substrate vial. Mix by gently vortexing, swirling or by inverting the contents to obtain a homogeneous solution. The CellTiter-Glo Substrate should go into solution easily in less than one minute.
[0096] MEK and ERK Inhibitor
[0097] MEK inhibitor Trametinib was purchased from Selleckchem (Cat No. 52673) and stored at -20.degree. C. before use. ERK1/2 inhibitor SCH772984 was purchased from Selleckchem (Cat No. 57101) and stored at -20.degree. C. before use.
[0098] Equipment
[0099] The following equipment was used in the experiments: EnVision Multi Label Reader 2104-0010A, PerkinElmer (USA); Countstar, Inno-Alliance Biotech (USA); Forma Series II Water Jacket CO2 Incubator, Thermo Scientific (USA); Biological safety Cabinet, Thermo Scientific, (USA); Inverted Microscope, Olympus CKX41SF (Japan).
[0100] Cytotoxicity and IC50 Determination
[0101] The day before the experiment (Day -1), cells were dissociated during the logarithmic growth period with Cell Disassociation Buffer (Gibco 13151-014) and mixed with appropriate cell media and centrifuge at 1000 rpm for 3 minutes. The cells were re-suspended and counted using Countstar before adjusting cell concentrations to optimized density (i.e. 4.44.times.10.sup.4 cells/ml) with respective culture medium listed in Table 1 for 3-day CTG assay (The cell density was optimized before actual study; cell density used in the test may vary for different cell lines). 90 .mu.l cell suspensions were added to two 96-well plates (plates A and B) with the final cell density of 4.times.10.sup.3 cells/well for 3-day CTG assay (the cell density was optimized before actual study; cell density used in the test may vary for different cell lines). The plate A and B group were incubated for overnight in humidified incubator at 37.degree. C. with 5% CO.sub.2.
[0102] On Day 0, for plate A group, 10 .mu.l culture medium was added to each well for T0 reading. CellTiter-Glo.RTM. Reagent was added at equal volume of cell culture medium present in each well (e.g., add 100 .mu.l of reagent to 100 .mu.l of medium containing cells for a 96-well plate). Contents were mixed for 2 minutes on an orbital shaker to facilitate cell lysis. The plate was allowed to incubate at room temperature for 10 minutes to stabilize luminescent signal. Backseal black sticker was added to the bottom of each plate. Luminescence was recorded using EnVision Multi Label Reader. This formed the basis for T0 value.
[0103] On Day 0, the test articles and positive controls were dissolved at the concentration indicated at Test Article Dilution map. 100.times. solution in PBS was prepared and then diluted with appropriate culture media (1:10) into 10.times.working solutions. 10 .mu.l (10.times.) drug solutions were dispensed in each well (triplicate for each drug concentration) of the plate B group according to plate inoculation map. The test plates were incubated for 4 days in the humidified incubator at 37.degree. with 5% CO.sub.2.
[0104] On Day 3, CellTiter-Glo.RTM. Reagent was added at equal volume of cell culture medium present in each well (e.g., add 100 .mu.l of reagent to 100 .mu.l of medium containing cells for a 96-well plate). Contents were mixed for 2 minutes on an orbital shaker to induce cell lysis. The plate was allowed to incubate at room temperature for 10 minutes to stabilize luminescent signal. Backseal black sticker was placed to the bottom of each plate. Luminescence was recorded using EnVision Multi Label Reader.
[0105] The data were displayed graphically using GraphPad Prism 5.0. In order to calculate IC50s, a dose-response curve was fitted using a nonlinear regression model with a sigmoidal dose response. The formula for calculating surviving rate was shown below; Absolute IC50 is calculated where Y axis set at 50% using GraphPad Prism 5.0. Software.
The surviving rate (%)=(Lum.sub.Test article-Lum.sub.Medium control)/(Lum.sub.None treated-Lum.sub.Medium control).times.100%.
[0106] Lum.sub.None treated-Lum.sub.Medium control is set as 100% and Lum.sub.Medium control is set for 0% surviving rate. T0 value was presented as percentage of Lum.sub.None treated.
[0107] Statistical Analysis
[0108] We divided the 63 cell lines into sensitive, insensitive, and uncertain groups according to their IC50's for SCH772984 and Trametinib, respectively, then detected genes with differential expression or different mutation types between sensitive and insensitive groups. These genes were enriched in several cancer related pathways.
[0109] The 63 cell lines were divided into 3 groups (See Table 1): a sensitive group (IC50 values were less than 1), an insensitive group (IC50 values are greater than 10), and an uncertain group (the rest). Accordingly, we got 25 sensitive and 22 insensitive cell lines for SCH772984, and 34 sensitive and 24 insensitive cell lines for Trametinib. Only cell lines with genomic data were used in subsequent analysis. The differentially expressed genes and enriched pathways were analysed using GSEA software, the genes with different mutation types were detected using Fisher's exact test.
[0110] Results
[0111] For Trametinib, 32 sensitive and 23 insensitive cell lines have gene expression profiled by Affymetrix U219 arrays, 34 gene sets are significantly enriched at nominal p-value <1% (See Table S2). 29 sensitive and 20 insensitive cell lines have mutation information, and ADAM12, COL14A1, TNN and TP53 were identified by P-value cutoff of 0.01 (See Table 3 and FIG. 1-5).
[0112] For SCH772984, 23 sensitive and 22 insensitive cell lines have gene expression profiled by Affymetrix U219 arrays, 10 gene sets are significantly enriched at nominal p-value <1%. 21 sensitive and 18 insensitive cell lines have mutation information, and ADAM12, PEX5L, TNN and TP53 were identified by P-value cutoff of 0.01 (See Table 3 and FIG. 6-10).
TABLE-US-00002 TABLE 2 IC50 information Absolute IC50 (uM) Number Cell line SCH772984 Trametinib 1 A549 1.78 0.24 2 A2058 NA 0.12 3 Calu6 0.56 0.14 4 DLD-1 52.77 6.52 5 HCT116 0.45 0.04 6 HepG2 0.13 0.00 7 MDA-MB-231 32.28 23.98 8 NCI-H23 3.69 0.30 9 NCI-H460 5.53 NA 10 RKO 17.46 6.70 11 SW620 0.26 0.01 12 SW480 NA 6.65 13 HCT-8 1.27 0.05 14 HCT-15 15.57 29.33 15 HT29 0.23 0.01 16 LoVo 0.41 0.11 17 LS513 0.78 0.01 18 NCI-H358 0.52 0.05 19 NCI-H441 NA 324.66 20 NCI-H1299 NA 0.91 21 NCI-H1792 4.70 0.16 22 PANC-1 64.31 330.05 23 Sk-Hep-1 5.29 29.14 24 Sk-Mel-5 0.20 0.01 25 BT474 NA 39.00 26 Colo205 0.04 0.00 27 KM12L4 0.36 0.01 28 MIAPaCa2 0.21 0.03 29 NCI-H1155 NA 636.34 30 NCI-H1373 NA NA 31 NCI-H1651 48.79 NA 32 NCI-H1666 0.92 0.13 33 NCI-H2009 4.36 0.41 34 NCI-H2227 28.74 NA 35 PL45 0.66 0.12 36 SK-LU-1 4.69 NA 37 SNU-1 0.64 0.10 38 ZR-75-1 NA NA 39 22RV1 5.01 202.02 40 BxPc-3 0.32 0.03 41 HCC2935 NA NA 42 HCC4006 2.77 0.41 43 Hela 5.38 NA 44 Hep3B 0.25 0.05 45 HM-7 0.79 0.39 46 HT1376 4.27 NA 47 KYSE150 1.72 5.28 48 NCI-H1703 NA 14.99 49 PC-3 14.29 NA 50 Du145 NA NA
TABLE-US-00003 TABLE 3 Gene Mutations Detected in MEK/ERK inhibitor insensitive cell lines. Variant Variant Tumor Gene Classification Type Sample Genome Change ADAM12 Missense SNP DU145 g.chr10: 127734680G > T ADAM12 Missense SNP DU145 g.chr10: 127797193C > A ADAM12 Missense SNP NCIH1573 g.chr10: 127760059C > T ADAM12 Missense SNP NCIH1573 g.chr10: 127797230G > C ADAM12 Missense SNP NCIH1703 g.chr10: 127797173G > C ADAM12 Missense SNP RKO g.chr10: 127787024C > T ADAM12 Frame Shift Del DEL RKO g.chr10: 127843846_127843846delT ADAM12 Missense SNP SW480 g.chr10: 127806716G > A ADAM12 Missense SNP HCT15 g.chr10: 127787067C > T COL14A1 Missense SNP 22RV1 g.chr8: 121209125C > T COL14A1 Splice Site SNP SNP DU145 g.chr8: 121239592G > T COL14A1 Missense SNP DU145 g.chr8: 121293288C > A COL14A1 Missense SNP NCIH1573 g.chr8: 121222108C > A COL14A1 Missense SNP NCIH2009 g.chr8: 121295933C > T COL14A1 Missense SNP NCIH441 g.chr8: 121279119G > A COL14A1 Missense SNP NCIH460 g.chr8: 121313055C > T COL14A1 Missense SNP RKO g.chr8: 121243717G > A COL14A1 Missense SNP SNU719 g.chr8: 121279117A > C COL14A1 Missense SNP HCT15 g.chr8: 121275133G > T COL14A1 Frame Shift Del DEL NCIH1373 g.chr8: 121326250_121326250delC TNN Missense SNP A549 g.chr1: 175052894G > A TNN Missense SNP DU145 g.chr1: 175049401A > C TNN Missense SNP NCIH1573 g.chr1: 175086152G > C TNN Missense SNP NCIH1703 g.chr1: 175067731G > T TNN Missense SNP NCIH2009 g.chr1: 175063212G > T TNN Missense SNP A2058 g.chr1: 175105993C > T TNN Missense SNP HCT15 g.chr1: 175063227C > T TNN Missense SNP HCT15 g.chr1: 175087926G > T TNN Missense SNP NCIH1299 g.chr1: 175063170G > T TNN Missense SNP NCIH1373 g.chr1: 175046753G > A TNN Missense SNP NCIH1373 g.chr1: 175087777A > T TNN Missense SNP NCIH2227 g.chr1: 175048841A > T TNN Missense SNP NCIH2227 g.chr1: 175087702G > T TNN Nonsense SNP NCIH2227 g.chr1: 175113638C > A TNN Missense SNP NCIH441 g.chr1: 175096204C > A TNN Missense SNP SKLU1 g.chr1: 175046766C > T TP53 Missense SNP 22RV1 g.chr17: 7576854T > C TP53 Frame Shift Del DEL ASPC1 g.chr17: 7578527_7578527delA TP53 Missense SNP BT474 g.chr17: 7577085C > T TP53 Missense SNP A2058 g.chr17: 7577118C > A TP53 Missense SNP BXPC3 g.chr17: 7578190T > C TP53 Nonsense SNP CALU6 g.chr17: 7578263G > A TP53 Missense SNP CAPAN1 g.chr17: 7578454G > A TP53 Missense SNP CFPAC1 g.chr17: 7577557A > G TP53 Missense SNP DU145 g.chr17: 7577118C > A TP53 Missense SNP HCC2935 g.chr17: 7578190T > C TP53 Missense SNP HT1376 g.chr17: 7577532G > A TP53 Missense SNP MDAMB231 g.chr17: 7577099C > T TP53 Missense SNP MIAPACA2 g.chr17: 7577539G > A TP53 Missense SNP NCIH1651 g.chr17: 7578403C > T TP53 Splice Site SNP SNP NCIH1703 g.chr17: 7577018C > A TP53 Splice Site SNP SNP NCIH1792 g.chr17: 7578176C > T TP53 Splice Site SNP SNP NCIH2227 g.chr17: 7577157T > G TP53 Missense SNP NCIH23 g.chr17: 7577543C > G TP53 Missense SNP NCIH441 g.chr17: 7578457C > A TP53 Frame Shift Del DEL PC3 g.chr17: 7578516_7578516delG TP53 Missense SNP SKLU1 g.chr17: 7578271T > C TP53 Nonsense SNP SNU387 g.chr17: 7578440T > A TP53 Missense SNP HUCCT1 g.chr17: 7578406C > T TP53 Missense SNP NCIH1573 g.chr17: 7577538C > A TP53 Missense SNP NCIH2009 g.chr17: 7577120C > A TP53 Missense SNP SW1116 g.chr17: 7578454G > T TP53 Missense SNP SW1271 g.chr17: 7577108C > A TP53 Missense SNP MIAPACA2 g.chr17: 7577539G > A PEX5L Missense SNP BT474 g.chr3: 179593236C > T PEX5L Missense SNP DU145 g.chr3: 179592155G > T PEX5L Missense SNP NCIH1573 g.chr3: 179754377C > T PEX5L Missense DNP NCIH2009 g.chr3: 179605504_179605505GG > TT PEX5L Missense SNP NCIH441 g.chr3: 179533669G > C PEX5L Missense SNP NCIH441 g.chr3: 179616013C > T PEX5L Missense SNP SW1271 g.chr3: 179519784C > A PEX5L Missense SNP NCIH1299 g.chr3: 179597885C > T Gene cDNA_Change Codon_Change Protein_Change ADAM12 c.1948C > A c.(1948-1950)CAA > AAA p.Q650K ADAM12 c.719G > T c.(718-720)CGA > CTA p.R240L ADAM12 c.1319G > A c.(1318-1320)TGT > TAT p.C440Y ADAM12 c.682C > G c.(682-684)CAG > GAG p.Q228E ADAM12 c.739C > G c.(739-741)CAC > GAC p.H247D ADAM12 c.966G > A c.(964-966)ATG > ATA p.M322I ADAM12 c.289_289delA c.(289-291)ACCfs p.T97fs ADAM12 c.503C > T c.(502-504)CCA > CTA p.P168L ADAM12 c.923G > A c.(922-924)GGG > GAG p.G308E COL14A1 c.532C > T c.(532-534)CGG > TGG p.R178W COL14A1 c.2137_splice p.L713_splice COL14A1 c.3814C > A c.(3814-3816)CAG > AAG p.Q1272K COL14A1 c.1435C > A c.(1435-1437)CTA > ATA p.L479I COL14A1 c.3883C > T c.(3883-3885)CTT > TTT p.L1295F COL14A1 c.3070G > A c.(3070-3072)GAA > AAA p.E1024K COL14A1 c.4399C > T c.(4399-4401)CCA > TCA p.P1467S COL14A1 c.2209G > A c.(2209-2211)GGA > AGA p.G737R COL14A1 c.3068A > C c.(3067-3069)AAA > ACA p.K1023T COL14A1 c.2896G > T c.(2896-2898)GGT > TGT p.G966C COL14A1 c.4535_4535delC c.(4534-4536)TCCfs p.S1512fs TNN c.1057G > A c.(1057-1059)GTG > ATG p.V353M TNN c.887A > C c.(886-888)TAC > TCC p.Y296S TNN c.2197G > C c.(2197-2199)GCC > CCC p.A733P TNN c.2119G > T c.(2119-2121)GAC > TAC p.D707Y TNN c.1411G > T c.(1411-1413)GAC > TAC p.D471Y TNN c.3464C > T c.(3463-3465)CCA > CTA p.P1155L TNN c.1426C > T c.(1426-1428)CGC > TGC p.R476C TNN c.2616G > T c.(2614-2616)CAG > CAT p.Q872H TNN c.1369G > T c.(1369-1371)GAC > TAC p.D457Y TNN c.199G > A c.(199-201)GAC > AAC p.D67N TNN c.2467A > T c.(2467-2469)ACC > TCC p.T823S TNN c.782A > T c.(781-783)CAG > CTG p.Q261L TNN c.2392G > T c.(2392-2394)GAC > TAC p.D798Y TNN c.3711C > A c.(3709-3711)TGC > TGA p.C1237* TNN c.3028C > A c.(3028-3030)CCA > ACA p.P1010T TNN c.212C > T c.(211-213)TCG > TTG p.S71L TP53 c.992A > G c.(991-993)CAG > CGG p.Q331R TP53 c.403_403delT c.(403-405)TGCfs p.C135fs TP53 c.853G > A c.(403-405)TGCfs pE285K TP53 c.820G > T c.(820-822)GTT > TTT p.V274F TP53 c.659A > G c.(658-660)TAT > TGT p.Y220C TP53 c.586C > T c.(586-588)CGA > TGA p.R196* TP53 c.476C > T c.(475-477)GCC > GTC p.A159V TP53 c.724T > C c.(724-726)TGC > CGC p.C242R TP53 c.820G > T c.(820-822)GTT > TTT p.V274F TP53 c.659A > G c.(658-660)TAT > TGT p.Y220C TP53 c.749C > T c.(748-750)CCC > CTC p.P250L TP53 c.839G > A c.(838-840)AGA > AAA p.R280K TP53 c.742C > T c.(742-744)CGG > TGG p.R248W TP53 c.527G > A c.(526-528)TGC > TAC p.C176Y TP53 c.919_splice c.e8 + 1 p.A307_splice TP53 c.672_splice c.e6 + 1 p.E224_splice TP53 c.783_splice c.e8 - 1 p.S261_splice TP53 c.738G > C c.(736-738)ATG > ATC p.M246I TP53 c.473G > T c.(472-474)CGC > CTC p.R158L TP53 c.414_414delC c.(412-414)GCCfs p.A138fs TP53 c.578A > G c.(577-579)CAT > CGT p.H193R TP53 c.490A > T c.(490-492)AAG > TAG p.K164* TP53 c.524G > A c.(523-525)CGC > CAC p.R175H TP53 c.743G > T c.(742-744)CGG > CTG p.R248L TP53 c.818G > T c.(817-819)CGT > CTT p.R273L TP53 c.476C > A c.(475-477)GCC > GAC p.A159D TP53 c.830G > T c.(829-831)TGT > TTT p.C277F TP53 c.742C > T c.(742-744)CGG > TGG p.R248W PEX5L c.535G > A c.(535-537)GAT > AAT p.D179N PEX5L c.686C > A c.(685-687)TCT > TAT p.S229Y PEX5L c.11G > A c.(10-12)GGA > GAA p.G4E PEX5L c.266_267CC > AA c.(265-267)ACC > AAA p.T89K PEX5L c.1063C < G c.(1063-1065)CAG > GAG p.Q355E PEX5L c.115G > A c.(115-117)GAT > AAT p.D39N PEX5L c.1713G > T c.(1711-1713)TTG > TTT p.L571F PEX5L c.337G > A c.(337-339)GAC > AAC p.D113N
[0113] While the invention has been particularly shown and described with reference to specific embodiments (some of which are preferred embodiments), it should be understood by those having skill in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the present invention as disclosed herein.
Sequence CWU
1
1
1018050DNAHomo sapiens 1acgccgccaa ctggcggggg tgcgggggag acaataattt
gttccgcggt aataagaacg 60gtgactgctg gccgtggatc catttcacag gcctgccttc
tctcactaac gctcttccta 120gtccccgggc caactcggac agtttgctca tttattgcaa
cggtcaaggc tggcttgtgc 180cagaacggcg cgcgcgcgcg cacgcacgca cacacacggg
gggaaacttt tttaaaaatg 240aaaggctaga agagctcagc ggcggcgcgg gcgctgcgcg
agggctccgg agctgactcg 300ccgaggcagg aaatccctcc ggtcgcgacg cccggccccg
gctcggcgcc cgcgtgggat 360ggtgcagcgc tcgccgccgg gcccgagagc tgctgcactg
aaggccggcg acgatggcag 420cgcgcccgct gcccgtgtcc cccgcccgcg ccctcctgct
cgccctggcc ggtgctctgc 480tcgcgccctg cgaggcccga ggggtgagct tatggaacca
aggaagagct gatgaagttg 540tcagtgcctc tgttgggagt ggggacctct ggatcccagt
gaagagcttc gactccaaga 600atcatccaga agtgctgaat attcgactac aacgggaaag
caaagaactg atcataaatc 660tggaaagaaa tgaaggtctc attgccagca gtttcacgga
aacccactat ctgcaagacg 720gtactgatgt ctccctcgct cgaaattaca cggtaattct
gggtcactgt tactaccatg 780gacatgtacg gggatattct gattcagcag tcagtctcag
cacgtgttct ggtctcaggg 840gacttattgt gtttgaaaat gaaagctatg tcttagaacc
aatgaaaagt gcaaccaaca 900gatacaaact cttcccagcg aagaagctga aaagcgtccg
gggatcatgt ggatcacatc 960acaacacacc aaacctcgct gcaaagaatg tgtttccacc
accctctcag acatgggcaa 1020gaaggcataa aagagagacc ctcaaggcaa ctaagtatgt
ggagctggtg atcgtggcag 1080acaaccgaga gtttcagagg caaggaaaag atctggaaaa
agttaagcag cgattaatag 1140agattgctaa tcacgttgac aagttttaca gaccactgaa
cattcggatc gtgttggtag 1200gcgtggaagt gtggaatgac atggacaaat gctctgtaag
tcaggaccca ttcaccagcc 1260tccatgaatt tctggactgg aggaagatga agcttctacc
tcgcaaatcc catgacaatg 1320cgcagcttgt cagtggggtt tatttccaag ggaccaccat
cggcatggcc ccaatcatga 1380gcatgtgcac ggcagaccag tctgggggaa ttgtcatgga
ccattcagac aatccccttg 1440gtgcagccgt gaccctggca catgagctgg gccacaattt
cgggatgaat catgacacac 1500tggacagggg ctgtagctgt caaatggcgg ttgagaaagg
aggctgcatc atgaacgctt 1560ccaccgggta cccatttccc atggtgttca gcagttgcag
caggaaggac ttggagacca 1620gcctggagaa aggaatgggg gtgtgcctgt ttaacctgcc
ggaagtcagg gagtctttcg 1680ggggccagaa gtgtgggaac agatttgtgg aagaaggaga
ggagtgtgac tgtggggagc 1740cagaggaatg tatgaatcgc tgctgcaatg ccaccacctg
taccctgaag ccggacgctg 1800tgtgcgcaca tgggctgtgc tgtgaagact gccagctgaa
gcctgcagga acagcgtgca 1860gggactccag caactcctgt gacctcccag agttctgcac
aggggccagc cctcactgcc 1920cagccaacgt gtacctgcac gatgggcact catgtcagga
tgtggacggc tactgctaca 1980atggcatctg ccagactcac gagcagcagt gtgtcacgct
ctggggacca ggtgctaaac 2040ctgcccctgg gatctgcttt gagagagtca attctgcagg
tgatccttat ggcaactgtg 2100gcaaagtctc gaagagttcc tttgccaaat gcgagatgag
agatgctaaa tgtggaaaaa 2160tccagtgtca aggaggtgcc agccggccag tcattggtac
caatgccgtt tccatagaaa 2220caaacatccc cctgcagcaa ggaggccgga ttctgtgccg
ggggacccac gtgtacttgg 2280gcgatgacat gccggaccca gggcttgtgc ttgcaggcac
aaagtgtgca gatggaaaaa 2340tctgcctgaa tcgtcaatgt caaaatatta gtgtctttgg
ggttcacgag tgtgcaatgc 2400agtgccacgg cagaggggtg tgcaacaaca ggaagaactg
ccactgcgag gcccactggg 2460cacctccctt ctgtgacaag tttggctttg gaggaagcac
agacagcggc cccatccggc 2520aagcagataa ccaaggttta accataggaa ttctggtgac
catcctgtgt cttcttgctg 2580ccggatttgt ggtttatctc aaaaggaaga ccttgatacg
actgctgttt acaaataaga 2640agaccaccat tgaaaaacta aggtgtgtgc gcccttcccg
gccaccccgt ggcttccaac 2700cctgtcaggc tcacctcggc caccttggaa aaggcctgat
gaggaagccg ccagattcct 2760acccaccgaa ggacaatccc aggagattgc tgcagtgtca
gaatgttgac atcagcagac 2820ccctcaacgg cctgaatgtc cctcagcccc agtcaactca
gcgagtgctt cctcccctcc 2880accgggctcc acgtgcacct agcgtccctg ccagacccct
gccagccaag cctgcactta 2940ggcaggccca ggggacctgt aagccaaacc cccctcagaa
gcctctgcct gcagatcctc 3000tggccagaac aactcggctc actcatgcct tggccaggac
cccaggacaa tgggagactg 3060ggctccgcct ggcacccctc agacctgctc cacaatatcc
acaccaagtg cccagatcca 3120cccacaccgc ctatattaag tgagaagccg acaccttttt
tcaacagtga agacagaagt 3180ttgcactatc tttcagctcc agttggagtt ttttgtacca
acttttagga ttttttttaa 3240tgtttaaaac atcattacta taagaacttt gagctactgc
cgtcagtgct gtgctgtgct 3300atggtgctct gtctacttgc tcaggtactt gtaaattatt
aatttatgca gaatgttgat 3360tacagtgcag tgcgctgtag taggcatttt taccatcact
gagttttcca tggcaggaag 3420gcttgttgtg cttttagtat tttagtgaac ttgaaatatc
ctgcttgatg ggattctgga 3480caggatgtgt ttgctttctg atcaaggcct tattggaaag
cagtccccca actaccccca 3540gctgtgctta tggtaccaga tgcagctcaa gagatcccaa
gtagaatctc agttgatttt 3600ctggattccc catctcaggc cagagccaag gggcttcagg
tccaggctgt gtttggcttt 3660cagggaggcc ctgtgcccct tgacaactgg caggcaggct
cccagggaca cctgggagaa 3720atctggcttc tggccaggaa gctttggtga gaacctgggt
tgcagacagg aatcttaagg 3780tgtagccaca ccaggataga gactggaaca ctagacaagc
cagaacttga ccctgagctg 3840accagccgtg agcatgtttg gaaggggtct gtagtgtcac
tcaaggcggt gcttgataga 3900aatgccaagc acttcttttt ctcgctgtcc tttctagagc
actgccacca gtaggttatt 3960tagcttggga aaggtggtgt ttctgtaaga aacctactgc
ccaggcactg caaaccgcca 4020cctccctata ctgcttggag ctgagcaaat caccacaaac
tgtaatacaa tgatcctgta 4080ttcagacaga tgaggctttc catgggacca caactatttt
cagatgtgaa ccattaacca 4140gatctagtca atcaagtctg tttactgcaa ggttcaactt
attaacaatt aggcagactc 4200tttatgcttg caaaaactac aaccaatgga atgtgatgtt
catgggtata gttcatgtct 4260gctatcatta ttcgtagata ttggacaaag aaccttctct
atggggcatc ctctttttcc 4320aacttggctg caggaatctt taaaagatgc ttttaacaga
gtctgaacct atttcttaaa 4380cacttgcaac ctacctgttg agcatcacag aatgtgataa
ggaaatcaac ttgcttatca 4440acttcctaaa tattatgaga tgctggcttg ggcagcatcc
ccttgaactc ttcactcttc 4500aaatgcctga ctagggagcc atgtttcaca aggtctttaa
agtgactaat ggcatgagaa 4560atacaaaaat actcagataa ggtaaaatgc catgatgcct
ctgtcttctg gactggtttt 4620cacattagaa gacaattgac aacagttaca taattcactc
tgagtgtttt atgagaaagc 4680cttcttttgg gggtcaacag ttttcctatg ctttgaaaca
gaaaaatatg taccaagaat 4740cttggtttgc cttccagaaa acaaaactgc atttcacttt
cccggtgttc cccactgtat 4800ctaggcaaca tagtattcat gactatggat aaactaaaca
cgtgacacaa acacacacaa 4860aagggaaccc agctctaata cattccaact cgtatagcat
gcatctgttt attctatagt 4920tattaagttc tttaaaatgt aaagccatgc tggaaaataa
tactgctgag atacatacag 4980aattactgta actgattaca cttggtaatt gtactaaagc
caaacatata tatactatta 5040aaaaggttta cagaatttta tggtgcatta cgtgggcatt
gtctttttag atgcccaaat 5100ccttagatct ggcatgttag cccttcctcc aattataaga
ggatatgaac tgagtttttc 5160ttttgttgtt tgttcttagc tgtaattcct atgcttctat
ttcagagagc caggagagtt 5220tgatattaaa ggaggttaaa actgtgatct tatgccatgt
catcaatggc cacttagggg 5280ccatggctga tgacacattc ttatctctac agtactaatg
tgttattata gagccatgca 5340ttttatttct gaataagaac atatttaaac taatattccc
ttacaatatg gacagtatta 5400atccttccaa gatgcagtat ttatcaagtg aagcatattt
agcagcaaat tccattttaa 5460cataacttag gaaccaataa ccagggtgtt ttgtggttgg
gggaggcacg gggtggagta 5520ttctttttta tatcctcaaa acaaaaaaaa tcaatactta
tatttcaatg gcaatctagt 5580atttttttaa aagactgtat aggcatgaat aatagaggtg
gtttgagttt tgtagggcca 5640tcacctggaa agtcaatgtg actagacaca aagtagccca
gaggctactt ttcttcctac 5700agcttattat agttgtaggt tctatgacct cacttcatgg
gttccaggca attccgctga 5760aaggtttgtc tcctgaaatt ttttaagttt gttttcctga
cacatgtaat cagatgtgta 5820gcaaccgagg gaaacgaagc ctaacattct ccattgtgga
aatacacaca ggaggttaca 5880tttcacagcg tggatttttc cagcttacac atgtgggatg
acatcacaga aaccacaaaa 5940gcagcaaatt aaactgtagg agagtcaata ctcctgacga
gtctcggggg gggggcattt 6000ttatgccttc ttaactttat gagaattctc aggctgaact
ataggccatt gttcccaggc 6060aaatcaatac atcaatgcat cctcaaaaaa aaaaaaaaaa
aaaaaaaacc ggctaaaact 6120gtgtcaaaat gttcttaagg agcctatggt ctccacggtg
ctaaaaagag cctggtgctg 6180ggccgactgg cagggctgag catcctcctg ccccctcgcc
actgatgttt actaagcact 6240ctgagccaat gagaccccca gcagcagaaa gggcacaagg
tggcgccagg gcagcagggc 6300cagatctttc tcatgcacct cgacctcttg cagactttct
tcgtgagatg tactactcat 6360ttcaaaactg ctttgcaggg ctcccctatg tattcggggg
gcccacggca cactcaggct 6420ggagatcctt cctcactgcg ctcaagatgg cctcagccag
acaccagtta cccagctgaa 6480agtcacaatc cctcccagaa gtctcccaac actagtgctg
accagaggtg gggctctcag 6540gctaggagtt tcacacacaa tgacaggctg ctgggggaca
ttgcaggacc ccttttcctc 6600tcctctccat gctagaagcc agccctaggc agctgcagtt
actccctgtg actcagcagc 6660aggctgattc aacacagctg cccacacaaa gccagtggct
aatacatctg tttacctttc 6720cctatcaccc agacacaagc ccctttccca ggtcaaacca
caggccgatg catctccagt 6780ttgacagtca aatcactact tccattgcta ctttagatca
gccaaagtgg tgactgctgc 6840agtgtgtggc tatccctaca aggcccaccc aagggatgcc
caaagcccaa ccttctccag 6900ggctgcagcc cagagcaacc ccaccagcct aagtccagca
gaggacctcc cacccaatgt 6960cttgttctaa ttagaagggg aagttagcca cagaaaatca
acttatctat aattacaaaa 7020ttctcttgac tcaccttaaa gttcctattg acatctactg
cttttaaacc tatttgaaaa 7080ctctgatact aaaacaaatg acactctaag aaagtttggg
agccccatgc tgagaaccat 7140ttctgtgcag tgaggatgtt tccagaagct acttacctac
atgtgaatgt gccattttct 7200ttccttttgt agagaaaatc ccctttactt tttggaacag
taatggcagc ttctagtaca 7260gccattacag tttcatatga gaaaaattaa gaataactat
aaaattgtta aaatatccaa 7320taatggataa tgatggccag aagatttaac atacaaagta
attctcaatg taaagctatt 7380cagctcttcc aggttgaatg ccctgtaacc caccctgacc
ttccacatca tcttcaaaaa 7440gcagtttctc tgttccccat gattctccta taaggtaact
ctttagtcct ccatttagca 7500cattttaaat cctccaaaga ataagtatca tgtgattatt
ttagctttac aaaaaaaaag 7560ttgaatggcg ttttattttc atggcctata agcaggtacc
ttagtagggc agatatagga 7620aaaacaaatt agagcaaaac aaatcctcta caaatccaag
gcaggaaaag tggtggcaga 7680gtgactcatt ctcctgtccc tcccatcagg tcaaatcagg
aggctgcagt gaatgcctgt 7740tctttgaatg tgtagcagtt gttcctgtaa ctctttaaaa
cttggctata ggctgtttag 7800cacagtacag attaaagata cagttacgta aacagcaaag
taattttata gtgcttcatc 7860catttatcat gctttggttt gctaattttt tcacatacct
ttttctatca cagtctgttg 7920cttttgtaca catttctcat attggggttc gacaggtaaa
cacaaactgc tatttcagta 7980gaaaaagtta ttgttatgaa tattaaaccc aataaattgt
ataaaggtaa atatcaaaaa 8040aaaaaaaaaa
80502909PRTHomo sapiens 2Met Ala Ala Arg Pro Leu
Pro Val Ser Pro Ala Arg Ala Leu Leu Leu1 5
10 15Ala Leu Ala Gly Ala Leu Leu Ala Pro Cys Glu Ala
Arg Gly Val Ser 20 25 30Leu
Trp Asn Gln Gly Arg Ala Asp Glu Val Val Ser Ala Ser Val Gly 35
40 45Ser Gly Asp Leu Trp Ile Pro Val Lys
Ser Phe Asp Ser Lys Asn His 50 55
60Pro Glu Val Leu Asn Ile Arg Leu Gln Arg Glu Ser Lys Glu Leu Ile65
70 75 80Ile Asn Leu Glu Arg
Asn Glu Gly Leu Ile Ala Ser Ser Phe Thr Glu 85
90 95Thr His Tyr Leu Gln Asp Gly Thr Asp Val Ser
Leu Ala Arg Asn Tyr 100 105
110Thr Val Ile Leu Gly His Cys Tyr Tyr His Gly His Val Arg Gly Tyr
115 120 125Ser Asp Ser Ala Val Ser Leu
Ser Thr Cys Ser Gly Leu Arg Gly Leu 130 135
140Ile Val Phe Glu Asn Glu Ser Tyr Val Leu Glu Pro Met Lys Ser
Ala145 150 155 160Thr Asn
Arg Tyr Lys Leu Phe Pro Ala Lys Lys Leu Lys Ser Val Arg
165 170 175Gly Ser Cys Gly Ser His His
Asn Thr Pro Asn Leu Ala Ala Lys Asn 180 185
190Val Phe Pro Pro Pro Ser Gln Thr Trp Ala Arg Arg His Lys
Arg Glu 195 200 205Thr Leu Lys Ala
Thr Lys Tyr Val Glu Leu Val Ile Val Ala Asp Asn 210
215 220Arg Glu Phe Gln Arg Gln Gly Lys Asp Leu Glu Lys
Val Lys Gln Arg225 230 235
240Leu Ile Glu Ile Ala Asn His Val Asp Lys Phe Tyr Arg Pro Leu Asn
245 250 255Ile Arg Ile Val Leu
Val Gly Val Glu Val Trp Asn Asp Met Asp Lys 260
265 270Cys Ser Val Ser Gln Asp Pro Phe Thr Ser Leu His
Glu Phe Leu Asp 275 280 285Trp Arg
Lys Met Lys Leu Leu Pro Arg Lys Ser His Asp Asn Ala Gln 290
295 300Leu Val Ser Gly Val Tyr Phe Gln Gly Thr Thr
Ile Gly Met Ala Pro305 310 315
320Ile Met Ser Met Cys Thr Ala Asp Gln Ser Gly Gly Ile Val Met Asp
325 330 335His Ser Asp Asn
Pro Leu Gly Ala Ala Val Thr Leu Ala His Glu Leu 340
345 350Gly His Asn Phe Gly Met Asn His Asp Thr Leu
Asp Arg Gly Cys Ser 355 360 365Cys
Gln Met Ala Val Glu Lys Gly Gly Cys Ile Met Asn Ala Ser Thr 370
375 380Gly Tyr Pro Phe Pro Met Val Phe Ser Ser
Cys Ser Arg Lys Asp Leu385 390 395
400Glu Thr Ser Leu Glu Lys Gly Met Gly Val Cys Leu Phe Asn Leu
Pro 405 410 415Glu Val Arg
Glu Ser Phe Gly Gly Gln Lys Cys Gly Asn Arg Phe Val 420
425 430Glu Glu Gly Glu Glu Cys Asp Cys Gly Glu
Pro Glu Glu Cys Met Asn 435 440
445Arg Cys Cys Asn Ala Thr Thr Cys Thr Leu Lys Pro Asp Ala Val Cys 450
455 460Ala His Gly Leu Cys Cys Glu Asp
Cys Gln Leu Lys Pro Ala Gly Thr465 470
475 480Ala Cys Arg Asp Ser Ser Asn Ser Cys Asp Leu Pro
Glu Phe Cys Thr 485 490
495Gly Ala Ser Pro His Cys Pro Ala Asn Val Tyr Leu His Asp Gly His
500 505 510Ser Cys Gln Asp Val Asp
Gly Tyr Cys Tyr Asn Gly Ile Cys Gln Thr 515 520
525His Glu Gln Gln Cys Val Thr Leu Trp Gly Pro Gly Ala Lys
Pro Ala 530 535 540Pro Gly Ile Cys Phe
Glu Arg Val Asn Ser Ala Gly Asp Pro Tyr Gly545 550
555 560Asn Cys Gly Lys Val Ser Lys Ser Ser Phe
Ala Lys Cys Glu Met Arg 565 570
575Asp Ala Lys Cys Gly Lys Ile Gln Cys Gln Gly Gly Ala Ser Arg Pro
580 585 590Val Ile Gly Thr Asn
Ala Val Ser Ile Glu Thr Asn Ile Pro Leu Gln 595
600 605Gln Gly Gly Arg Ile Leu Cys Arg Gly Thr His Val
Tyr Leu Gly Asp 610 615 620Asp Met Pro
Asp Pro Gly Leu Val Leu Ala Gly Thr Lys Cys Ala Asp625
630 635 640Gly Lys Ile Cys Leu Asn Arg
Gln Cys Gln Asn Ile Ser Val Phe Gly 645
650 655Val His Glu Cys Ala Met Gln Cys His Gly Arg Gly
Val Cys Asn Asn 660 665 670Arg
Lys Asn Cys His Cys Glu Ala His Trp Ala Pro Pro Phe Cys Asp 675
680 685Lys Phe Gly Phe Gly Gly Ser Thr Asp
Ser Gly Pro Ile Arg Gln Ala 690 695
700Asp Asn Gln Gly Leu Thr Ile Gly Ile Leu Val Thr Ile Leu Cys Leu705
710 715 720Leu Ala Ala Gly
Phe Val Val Tyr Leu Lys Arg Lys Thr Leu Ile Arg 725
730 735Leu Leu Phe Thr Asn Lys Lys Thr Thr Ile
Glu Lys Leu Arg Cys Val 740 745
750Arg Pro Ser Arg Pro Pro Arg Gly Phe Gln Pro Cys Gln Ala His Leu
755 760 765Gly His Leu Gly Lys Gly Leu
Met Arg Lys Pro Pro Asp Ser Tyr Pro 770 775
780Pro Lys Asp Asn Pro Arg Arg Leu Leu Gln Cys Gln Asn Val Asp
Ile785 790 795 800Ser Arg
Pro Leu Asn Gly Leu Asn Val Pro Gln Pro Gln Ser Thr Gln
805 810 815Arg Val Leu Pro Pro Leu His
Arg Ala Pro Arg Ala Pro Ser Val Pro 820 825
830Ala Arg Pro Leu Pro Ala Lys Pro Ala Leu Arg Gln Ala Gln
Gly Thr 835 840 845Cys Lys Pro Asn
Pro Pro Gln Lys Pro Leu Pro Ala Asp Pro Leu Ala 850
855 860Arg Thr Thr Arg Leu Thr His Ala Leu Ala Arg Thr
Pro Gly Gln Trp865 870 875
880Glu Thr Gly Leu Arg Leu Ala Pro Leu Arg Pro Ala Pro Gln Tyr Pro
885 890 895His Gln Val Pro Arg
Ser Thr His Thr Ala Tyr Ile Lys 900
90536464DNAHomo sapiens 3atctgatcct gggctcccag ctggagaggc ggaggcagct
ccagggggct ggaagtggaa 60gcgcagcggc agaaggagag ggagagagaa agagagagag
gctaattaaa aaaggatact 120ccgagggaag agagcaaggg cggtgcgccg ccaaggacca
actagcggcg gagcttcgat 180cttgcctagg cgcggagagc tcccaacctg ggctggaacc
ttgcccagca caggtggctg 240ctacacccca tgtaaaaagc ggaaaataaa atgaagattt
tccagcgcaa gatgcggtac 300tggttgcttc cacctttttt ggcaattgtt tatttctgca
ccattgtcca aggtcaagtg 360gctccaccca caaggttaag atataatgta atatctcatg
acagtataca gatttcatgg 420aaggctccaa gagggaaatt tggtggttac aaacttcttg
tgactccaac ttcaggtgga 480aaaactaacc agctgaatct gcagaacact gcaactaaag
caattattca aggccttatg 540ccagaccaga attacacagt tcaaattatt gcatacaata
aagataaaga aagcaagcca 600gctcaaggcc aattcagaat taaagattta gaaaaaagaa
aggatccaaa gcccagagtc 660aaagttgtgg acagaggaaa tgggagtaga ccatcttcac
cagaagaagt gaaatttgtc 720tgtcaaactc cagcaattgc tgacattgta atcctggtcg
atggttcatg gagtattgga 780agattcaact tcagactggt tcggcatttc ttggaaaacc
tggttacagc attcgatgtg 840ggctcagaga agacacgaat tggtcttgca cagtatagtg
gtgaccccag aatagaatgg 900cacttgaatg catttagcac aaaagatgaa gtgattgaag
ctgtccgaaa cctcccatat 960aaaggaggaa atacactaac aggtcttgct ttgaactaca
tttttgaaaa tagcttcaaa 1020ccagaagcag gatcaaggac tggagtatcc aaaattggca
ttttaatcac agatggaaaa 1080tcccaagatg acattattcc accatctaga aatcttcgtg
agtctggtgt agaactgttt 1140gccatagggg tgaaaaacgc ggatgtgaat gagctgcagg
agatcgcctc tgaaccagac 1200agcactcatg tgtacaatgt tgccgaattc gatctgatgc
acacagttgt ggagagtctg 1260accaggactc tctgctctag agtggaagaa caggacagag
aaattaaagc ctcagcccat 1320gccatcactg ggccgcctac ggagttgatt acttctgaag
tcactgccag aagctttatg 1380gttaactgga ctcatgcccc aggaaatgtg gaaaaataca
gagttgtgta ttatcctacc 1440aggggtggaa aaccagacga ggtggtggta gatggaactg
tatcttccac agtgttgaaa 1500aacttgatgt ctttaactga atatcagata gcagtctttg
caatctatgc ccacactgct 1560agtgaaggcc tacggggaac tgaaactaca cttgctttac
cgatggcttc tgaccttcta 1620ctgtacgacg tgactgagaa cagcatgcga gtcaaatggg
atgcagtgcc tggggcctca 1680ggttacctga tcctttatgc tcctctaaca gagggcctgg
ctggggatga aaaagagatg 1740aaaattggag agacccacac agatattgaa ttgagtgggt
tgttgcccaa tacagaatac 1800acagtcacag tttatgccat gtttggagaa gaggccagtg
atcctgttac gggacaagaa 1860acaacattgg ctttaagtcc accaagaaac ctgagaatct
ccaatgttgg ctctaacagt 1920gctcgattaa cctgggaccc aacttcaaga cagatcaatg
gttatcgaat tgtatataac 1980aatgcagatg ggactgaaat caatgaggtt gaagtcgatc
ctattactac cttccctctg 2040aagggcttga cacctctcac agagtatact attgctattt
tctccatcta tgatgaagga 2100cagtcagagc ctctgactgg agtttttacc accgaggaag
ttccagccca gcaatactta 2160gaaattgatg aggtgacgac agacagtttt agggtgacct
ggcatcccct ctcagctgat 2220gaagggctac acaaattgat gtggattcca gtctatgggg
ggaagactga ggaggttgtc 2280ctgaaagaag agcaggactc acatgttatt gaaggcctgg
agcccggtac ggagtatgaa 2340gtttcactat tggccgtact tgatgatgga agcgagagtg
aggtggtgac tgctgtcggg 2400accacacttg acagtttttg gacagaacca gctacaacca
tagtgcctac cacatctgtg 2460acttcagttt tccagacggg aatcagaaac ctagttgtag
gtgatgaaac tacttctagc 2520ctgcgggtaa aatgggacat ttctgacagc gatgtgcagc
agtttagggt gacctacatg 2580acagctcaag gggaccctga ggaagaagtc ataggaacgg
ttatggtgcc tggaagccag 2640aacaacctcc ttctgaagcc tctgcttcct gatactgaat
acaaagtcac agtgactccc 2700atctacacgg atggcgaagg cgtcagcgtc tccgctcctg
gaaaaacctt accatcctcg 2760gggccccaga acttgcgggt gtccgaggaa tggtataacc
ggttgcgcat tacgtgggac 2820cccccatctt ccccggtgaa aggctataga attgtctaca
aacctgtcag tgttcctggt 2880ccaacactgg aaacgtttgt gggagctgac attaacacca
tccttatcac aaacctcctc 2940agcggaatgg actacaatgt gaagatattt gcctcccagg
cctcaggctt cagcgacgcc 3000ctgacaggca tggtgaaaac attgttcttg ggtgttacca
atctccaagc caaacatgtt 3060gaaatgacca gcttgtgtgc ccactggcag gtacatcgcc
atgccacagc ctatagggtt 3120gttatagaat ccctccagga taggcaaaag caagaatcca
ctgtgggtgg agggacaacc 3180aggcattgct tctatggact tcagcctgat tctgaatata
aaatcagtgt ttatacaaag 3240ctccaggaga ttgaaggacc tagtgtgagc ataatggaaa
aaacacaatc acttcctaca 3300cgaccaccaa cttttcctcc aaccattcca ccagcaaaag
aagtatgtaa ggcggccaag 3360gctgacctgg tatttatggt ggatggatcc tggagcattg
gagatgaaaa tttcaataag 3420atcatcagct ttctatacag cactgttgga gccctgaaca
agattggcac agatggaacc 3480caagttgcaa tggttcagtt cactgatgat cccagaacag
aatttaaact aaatgcttac 3540aaaaccaaag agactcttct tgatgcaatt aaacacattt
catacaaagg aggaaataca 3600aaaacaggaa aagcaattaa gtatgttcga gataccttgt
tcactgcaga gtcaggtaca 3660agaaggggca tcccaaaggt tatcgtggtt ataactgatg
gaagatcaca agatgatgtg 3720aacaaaatct ccagggagat gcaattagat ggctatagca
tttttgcaat tggtgtggcc 3780gatgcagatt actcggagtt ggttagcatt ggcagtaagc
ccagcgcacg ccatgtcttc 3840tttgtggatg actttgacgc ctttaagaaa atcgaagatg
agttaattac ttttgtctgc 3900gaaacagcat cagcaacctg tccagtggta cacaaggatg
gcattgatct tgcaggattt 3960aagatgatgg aaatgtttgg tttggttgaa aaagattttt
catcagtgga aggggtttct 4020atggagcctg gtaccttcaa tgtgtttcca tgttaccaac
tccataaaga tgccctggtt 4080tcccagccaa ccaggtactt gcacccagaa ggattgccct
ccgactacac aatcagtttt 4140ctattccgga ttcttcctga cactccacag gagccatttg
ctctttggga gattttaaat 4200aaaaattctg acccattggt tggggttatt ttagacaatg
gtgggaaaac tctaacatat 4260ttcaactatg accagagtgg ggattttcaa actgttactt
tcgaaggacc tgaaattagg 4320aaaatttttt atggaagctt tcacaagcta cacattgttg
tcagtgagac tttggtcaaa 4380gtggttattg actgcaagca agtgggtgag aaggcaatga
acgcatcagc taatatcacg 4440tcagatggtg tagaagtgct agggaaaatg gttcgatcaa
gaggaccagg tggaaactct 4500gcaccgttcc agttacagat gtttgatatt gtttgctcca
catcatgggc caatacagac 4560aaatgctgtg aacttccagg cctgagagat gatgagtctt
gcccagacct tccccattcc 4620tgctcctgtt ctgaaaccaa tgaagtggct ctgggaccag
cgggcccacc aggtggtcca 4680ggactccgag gaccaaaggg ccagcaaggt gaaccgggtc
caaagggacc agatggccct 4740cggggtgaaa ttggtctgcc aggacctcag ggtccacctg
gacctcaagg accaagtggt 4800ctgtccattc aaggaatgcc cggaatgcca ggagaaaaag
gagagaaagg agatactggc 4860cttccaggtc cacagggtat cccaggaggc gttggttcac
caggacgtga tggctcacca 4920ggccagaggg gccttccggg aaaggatgga tcctcgggac
ctccaggacc accagggcca 4980ataggcattc ctggcacccc tggagtccca gggatcacag
gaagcatggg accgcaaggc 5040gccctgggac cacctggtgt ccctggagca aagggggaac
gaggagagcg gggtgacctg 5100cagtctcaag ccatggtgag atcagtggcg cgtcaagtat
gcgaacagct catccagagt 5160cacatggcca ggtacactgc catcctcaac cagattccca
gccactcctc atccatccgg 5220actgtccaag ggcctcctgg ggagcctggg aggccaggct
cacctggagc ccctggtgaa 5280caaggacccc caggcacacc aggcttcccc ggaaatgcag
gcgtgccagg gaccccagga 5340gaacgaggtc taactggtat caaaggagaa aaaggaaatc
caggcgttgg aacccaaggt 5400ccaagaggcc cccctggacc agcaggacct tcaggggaga
gtcggcctgg cagccctggg 5460ccccctggct ctcctggacc aagaggccca ccaggtcatc
tgggggttcc tggaccccaa 5520ggtccttctg gccagcctgg atattgtgac ccctcatcat
gttctgccta tggtgtgaga 5580gctccccatc cagatcagcc agagttcacc cctgtccaag
atgagctgga agccatggaa 5640ctgtggggcc ctggagtctg atagcctcag gagaaatttg
aagaccaact gcaagaactc 5700ttaaggaatc ttgtttgaga aaatgttgtt atgtggtttg
tatgctactt ttggggggca 5760gggctcattt cagcagccta aatctcctcc ttggataatg
ttaatattat tattattatt 5820aacaaaaaat atatattttt aaaaagttcc cttaatctat
gacatggtag caatgatttc 5880cctttggtgt cttaatggca tgtcagataa tttgtttttc
cagagaagag agctcaaaga 5940ggaattggga aaaataaatt gaactctgga atcttctctc
tcaagtccta aaatgaacaa 6000acagatatga ttgtgtttga gggaaatatg tccctagcag
gaaaagaatt caaagaggtt 6060caaagaatat gtcacttact cctacttgct gtaggaataa
ccttgctgat aagaaaaaaa 6120gggacaatat tggagaaact acctcttgtt taattgatct
gtccaactct gagatcactt 6180ggtaactggt ttcatgtgta tccaaaaatc agcatttgga
tttaagcttt ctgaatttgg 6240tagtttaaga aacagattta gtttttcagt ggttttaact
catgtgaaat aatgattttc 6300caccagctct gatgcaaaga gatataattt taatgaacga
tttatccagc agtgtgttcc 6360aggggttgcc tctccttatc tacggggatt actttgtaca
tgcagataag ttttcgcaaa 6420cctatttcca ttttcttttg taagcaaata aaactttaaa
acaa 646441796PRTHomo sapiens 4Met Lys Ile Phe Gln Arg
Lys Met Arg Tyr Trp Leu Leu Pro Pro Phe1 5
10 15Leu Ala Ile Val Tyr Phe Cys Thr Ile Val Gln Gly
Gln Val Ala Pro 20 25 30Pro
Thr Arg Leu Arg Tyr Asn Val Ile Ser His Asp Ser Ile Gln Ile 35
40 45Ser Trp Lys Ala Pro Arg Gly Lys Phe
Gly Gly Tyr Lys Leu Leu Val 50 55
60Thr Pro Thr Ser Gly Gly Lys Thr Asn Gln Leu Asn Leu Gln Asn Thr65
70 75 80Ala Thr Lys Ala Ile
Ile Gln Gly Leu Met Pro Asp Gln Asn Tyr Thr 85
90 95Val Gln Ile Ile Ala Tyr Asn Lys Asp Lys Glu
Ser Lys Pro Ala Gln 100 105
110Gly Gln Phe Arg Ile Lys Asp Leu Glu Lys Arg Lys Asp Pro Lys Pro
115 120 125Arg Val Lys Val Val Asp Arg
Gly Asn Gly Ser Arg Pro Ser Ser Pro 130 135
140Glu Glu Val Lys Phe Val Cys Gln Thr Pro Ala Ile Ala Asp Ile
Val145 150 155 160Ile Leu
Val Asp Gly Ser Trp Ser Ile Gly Arg Phe Asn Phe Arg Leu
165 170 175Val Arg His Phe Leu Glu Asn
Leu Val Thr Ala Phe Asp Val Gly Ser 180 185
190Glu Lys Thr Arg Ile Gly Leu Ala Gln Tyr Ser Gly Asp Pro
Arg Ile 195 200 205Glu Trp His Leu
Asn Ala Phe Ser Thr Lys Asp Glu Val Ile Glu Ala 210
215 220Val Arg Asn Leu Pro Tyr Lys Gly Gly Asn Thr Leu
Thr Gly Leu Ala225 230 235
240Leu Asn Tyr Ile Phe Glu Asn Ser Phe Lys Pro Glu Ala Gly Ser Arg
245 250 255Thr Gly Val Ser Lys
Ile Gly Ile Leu Ile Thr Asp Gly Lys Ser Gln 260
265 270Asp Asp Ile Ile Pro Pro Ser Arg Asn Leu Arg Glu
Ser Gly Val Glu 275 280 285Leu Phe
Ala Ile Gly Val Lys Asn Ala Asp Val Asn Glu Leu Gln Glu 290
295 300Ile Ala Ser Glu Pro Asp Ser Thr His Val Tyr
Asn Val Ala Glu Phe305 310 315
320Asp Leu Met His Thr Val Val Glu Ser Leu Thr Arg Thr Leu Cys Ser
325 330 335Arg Val Glu Glu
Gln Asp Arg Glu Ile Lys Ala Ser Ala His Ala Ile 340
345 350Thr Gly Pro Pro Thr Glu Leu Ile Thr Ser Glu
Val Thr Ala Arg Ser 355 360 365Phe
Met Val Asn Trp Thr His Ala Pro Gly Asn Val Glu Lys Tyr Arg 370
375 380Val Val Tyr Tyr Pro Thr Arg Gly Gly Lys
Pro Asp Glu Val Val Val385 390 395
400Asp Gly Thr Val Ser Ser Thr Val Leu Lys Asn Leu Met Ser Leu
Thr 405 410 415Glu Tyr Gln
Ile Ala Val Phe Ala Ile Tyr Ala His Thr Ala Ser Glu 420
425 430Gly Leu Arg Gly Thr Glu Thr Thr Leu Ala
Leu Pro Met Ala Ser Asp 435 440
445Leu Leu Leu Tyr Asp Val Thr Glu Asn Ser Met Arg Val Lys Trp Asp 450
455 460Ala Val Pro Gly Ala Ser Gly Tyr
Leu Ile Leu Tyr Ala Pro Leu Thr465 470
475 480Glu Gly Leu Ala Gly Asp Glu Lys Glu Met Lys Ile
Gly Glu Thr His 485 490
495Thr Asp Ile Glu Leu Ser Gly Leu Leu Pro Asn Thr Glu Tyr Thr Val
500 505 510Thr Val Tyr Ala Met Phe
Gly Glu Glu Ala Ser Asp Pro Val Thr Gly 515 520
525Gln Glu Thr Thr Leu Ala Leu Ser Pro Pro Arg Asn Leu Arg
Ile Ser 530 535 540Asn Val Gly Ser Asn
Ser Ala Arg Leu Thr Trp Asp Pro Thr Ser Arg545 550
555 560Gln Ile Asn Gly Tyr Arg Ile Val Tyr Asn
Asn Ala Asp Gly Thr Glu 565 570
575Ile Asn Glu Val Glu Val Asp Pro Ile Thr Thr Phe Pro Leu Lys Gly
580 585 590Leu Thr Pro Leu Thr
Glu Tyr Thr Ile Ala Ile Phe Ser Ile Tyr Asp 595
600 605Glu Gly Gln Ser Glu Pro Leu Thr Gly Val Phe Thr
Thr Glu Glu Val 610 615 620Pro Ala Gln
Gln Tyr Leu Glu Ile Asp Glu Val Thr Thr Asp Ser Phe625
630 635 640Arg Val Thr Trp His Pro Leu
Ser Ala Asp Glu Gly Leu His Lys Leu 645
650 655Met Trp Ile Pro Val Tyr Gly Gly Lys Thr Glu Glu
Val Val Leu Lys 660 665 670Glu
Glu Gln Asp Ser His Val Ile Glu Gly Leu Glu Pro Gly Thr Glu 675
680 685Tyr Glu Val Ser Leu Leu Ala Val Leu
Asp Asp Gly Ser Glu Ser Glu 690 695
700Val Val Thr Ala Val Gly Thr Thr Leu Asp Ser Phe Trp Thr Glu Pro705
710 715 720Ala Thr Thr Ile
Val Pro Thr Thr Ser Val Thr Ser Val Phe Gln Thr 725
730 735Gly Ile Arg Asn Leu Val Val Gly Asp Glu
Thr Thr Ser Ser Leu Arg 740 745
750Val Lys Trp Asp Ile Ser Asp Ser Asp Val Gln Gln Phe Arg Val Thr
755 760 765Tyr Met Thr Ala Gln Gly Asp
Pro Glu Glu Glu Val Ile Gly Thr Val 770 775
780Met Val Pro Gly Ser Gln Asn Asn Leu Leu Leu Lys Pro Leu Leu
Pro785 790 795 800Asp Thr
Glu Tyr Lys Val Thr Val Thr Pro Ile Tyr Thr Asp Gly Glu
805 810 815Gly Val Ser Val Ser Ala Pro
Gly Lys Thr Leu Pro Ser Ser Gly Pro 820 825
830Gln Asn Leu Arg Val Ser Glu Glu Trp Tyr Asn Arg Leu Arg
Ile Thr 835 840 845Trp Asp Pro Pro
Ser Ser Pro Val Lys Gly Tyr Arg Ile Val Tyr Lys 850
855 860Pro Val Ser Val Pro Gly Pro Thr Leu Glu Thr Phe
Val Gly Ala Asp865 870 875
880Ile Asn Thr Ile Leu Ile Thr Asn Leu Leu Ser Gly Met Asp Tyr Asn
885 890 895Val Lys Ile Phe Ala
Ser Gln Ala Ser Gly Phe Ser Asp Ala Leu Thr 900
905 910Gly Met Val Lys Thr Leu Phe Leu Gly Val Thr Asn
Leu Gln Ala Lys 915 920 925His Val
Glu Met Thr Ser Leu Cys Ala His Trp Gln Val His Arg His 930
935 940Ala Thr Ala Tyr Arg Val Val Ile Glu Ser Leu
Gln Asp Arg Gln Lys945 950 955
960Gln Glu Ser Thr Val Gly Gly Gly Thr Thr Arg His Cys Phe Tyr Gly
965 970 975Leu Gln Pro Asp
Ser Glu Tyr Lys Ile Ser Val Tyr Thr Lys Leu Gln 980
985 990Glu Ile Glu Gly Pro Ser Val Ser Ile Met Glu
Lys Thr Gln Ser Leu 995 1000
1005Pro Thr Arg Pro Pro Thr Phe Pro Pro Thr Ile Pro Pro Ala Lys
1010 1015 1020Glu Val Cys Lys Ala Ala
Lys Ala Asp Leu Val Phe Met Val Asp 1025 1030
1035Gly Ser Trp Ser Ile Gly Asp Glu Asn Phe Asn Lys Ile Ile
Ser 1040 1045 1050Phe Leu Tyr Ser Thr
Val Gly Ala Leu Asn Lys Ile Gly Thr Asp 1055 1060
1065Gly Thr Gln Val Ala Met Val Gln Phe Thr Asp Asp Pro
Arg Thr 1070 1075 1080Glu Phe Lys Leu
Asn Ala Tyr Lys Thr Lys Glu Thr Leu Leu Asp 1085
1090 1095Ala Ile Lys His Ile Ser Tyr Lys Gly Gly Asn
Thr Lys Thr Gly 1100 1105 1110Lys Ala
Ile Lys Tyr Val Arg Asp Thr Leu Phe Thr Ala Glu Ser 1115
1120 1125Gly Thr Arg Arg Gly Ile Pro Lys Val Ile
Val Val Ile Thr Asp 1130 1135 1140Gly
Arg Ser Gln Asp Asp Val Asn Lys Ile Ser Arg Glu Met Gln 1145
1150 1155Leu Asp Gly Tyr Ser Ile Phe Ala Ile
Gly Val Ala Asp Ala Asp 1160 1165
1170Tyr Ser Glu Leu Val Ser Ile Gly Ser Lys Pro Ser Ala Arg His
1175 1180 1185Val Phe Phe Val Asp Asp
Phe Asp Ala Phe Lys Lys Ile Glu Asp 1190 1195
1200Glu Leu Ile Thr Phe Val Cys Glu Thr Ala Ser Ala Thr Cys
Pro 1205 1210 1215Val Val His Lys Asp
Gly Ile Asp Leu Ala Gly Phe Lys Met Met 1220 1225
1230Glu Met Phe Gly Leu Val Glu Lys Asp Phe Ser Ser Val
Glu Gly 1235 1240 1245Val Ser Met Glu
Pro Gly Thr Phe Asn Val Phe Pro Cys Tyr Gln 1250
1255 1260Leu His Lys Asp Ala Leu Val Ser Gln Pro Thr
Arg Tyr Leu His 1265 1270 1275Pro Glu
Gly Leu Pro Ser Asp Tyr Thr Ile Ser Phe Leu Phe Arg 1280
1285 1290Ile Leu Pro Asp Thr Pro Gln Glu Pro Phe
Ala Leu Trp Glu Ile 1295 1300 1305Leu
Asn Lys Asn Ser Asp Pro Leu Val Gly Val Ile Leu Asp Asn 1310
1315 1320Gly Gly Lys Thr Leu Thr Tyr Phe Asn
Tyr Asp Gln Ser Gly Asp 1325 1330
1335Phe Gln Thr Val Thr Phe Glu Gly Pro Glu Ile Arg Lys Ile Phe
1340 1345 1350Tyr Gly Ser Phe His Lys
Leu His Ile Val Val Ser Glu Thr Leu 1355 1360
1365Val Lys Val Val Ile Asp Cys Lys Gln Val Gly Glu Lys Ala
Met 1370 1375 1380Asn Ala Ser Ala Asn
Ile Thr Ser Asp Gly Val Glu Val Leu Gly 1385 1390
1395Lys Met Val Arg Ser Arg Gly Pro Gly Gly Asn Ser Ala
Pro Phe 1400 1405 1410Gln Leu Gln Met
Phe Asp Ile Val Cys Ser Thr Ser Trp Ala Asn 1415
1420 1425Thr Asp Lys Cys Cys Glu Leu Pro Gly Leu Arg
Asp Asp Glu Ser 1430 1435 1440Cys Pro
Asp Leu Pro His Ser Cys Ser Cys Ser Glu Thr Asn Glu 1445
1450 1455Val Ala Leu Gly Pro Ala Gly Pro Pro Gly
Gly Pro Gly Leu Arg 1460 1465 1470Gly
Pro Lys Gly Gln Gln Gly Glu Pro Gly Pro Lys Gly Pro Asp 1475
1480 1485Gly Pro Arg Gly Glu Ile Gly Leu Pro
Gly Pro Gln Gly Pro Pro 1490 1495
1500Gly Pro Gln Gly Pro Ser Gly Leu Ser Ile Gln Gly Met Pro Gly
1505 1510 1515Met Pro Gly Glu Lys Gly
Glu Lys Gly Asp Thr Gly Leu Pro Gly 1520 1525
1530Pro Gln Gly Ile Pro Gly Gly Val Gly Ser Pro Gly Arg Asp
Gly 1535 1540 1545Ser Pro Gly Gln Arg
Gly Leu Pro Gly Lys Asp Gly Ser Ser Gly 1550 1555
1560Pro Pro Gly Pro Pro Gly Pro Ile Gly Ile Pro Gly Thr
Pro Gly 1565 1570 1575Val Pro Gly Ile
Thr Gly Ser Met Gly Pro Gln Gly Ala Leu Gly 1580
1585 1590Pro Pro Gly Val Pro Gly Ala Lys Gly Glu Arg
Gly Glu Arg Gly 1595 1600 1605Asp Leu
Gln Ser Gln Ala Met Val Arg Ser Val Ala Arg Gln Val 1610
1615 1620Cys Glu Gln Leu Ile Gln Ser His Met Ala
Arg Tyr Thr Ala Ile 1625 1630 1635Leu
Asn Gln Ile Pro Ser His Ser Ser Ser Ile Arg Thr Val Gln 1640
1645 1650Gly Pro Pro Gly Glu Pro Gly Arg Pro
Gly Ser Pro Gly Ala Pro 1655 1660
1665Gly Glu Gln Gly Pro Pro Gly Thr Pro Gly Phe Pro Gly Asn Ala
1670 1675 1680Gly Val Pro Gly Thr Pro
Gly Glu Arg Gly Leu Thr Gly Ile Lys 1685 1690
1695Gly Glu Lys Gly Asn Pro Gly Val Gly Thr Gln Gly Pro Arg
Gly 1700 1705 1710Pro Pro Gly Pro Ala
Gly Pro Ser Gly Glu Ser Arg Pro Gly Ser 1715 1720
1725Pro Gly Pro Pro Gly Ser Pro Gly Pro Arg Gly Pro Pro
Gly His 1730 1735 1740Leu Gly Val Pro
Gly Pro Gln Gly Pro Ser Gly Gln Pro Gly Tyr 1745
1750 1755Cys Asp Pro Ser Ser Cys Ser Ala Tyr Gly Val
Arg Ala Pro His 1760 1765 1770Pro Asp
Gln Pro Glu Phe Thr Pro Val Gln Asp Glu Leu Glu Ala 1775
1780 1785Met Glu Leu Trp Gly Pro Gly Val 1790
179555008DNAHomo sapiens 5aagtaccaag gtctgcggca ggaggagacc
ggctcacagg agcagcagca ttggaagagg 60cacccagcag cctcccaggc atcctggagg
gtctgctccc tgtctttcca aggatgagtc 120tccaggagat gttccgcttc cctatggggc
tcctgcttgg ctctgtgctc ctggtggctt 180cggccccagc cactctggag cctcccggct
gcagcaacaa ggagcaacag gtcactgtca 240gccacaccta caagatcgat gtgcccaagt
ctgccttggt tcaggttgac gctgaccctc 300agcccctcag tgacgatggg gcttcgctct
tggccctggg ggaggccagg gaggaacaga 360acatcatctt caggcacaac atccgccttc
agacgccaca gaaggactgc gagttggcag 420gcagtgtcca ggacctcctg gcccgggtga
agaagctgga ggaagagatg gtggagatga 480aggaacagtg tagtgcccag cgctgctgcc
agggagtcac tgatctaagc cgccactgca 540gcggccacgg gaccttctcc ctggagacct
gcagctgcca ctgcgaagag ggcagggagg 600gccccgcctg cgagcggctg gcctgccccg
gggcgtgcag cggccacggg cgttgcgtgg 660acgggcgctg cctgtgccat gagccctacg
tgggtgccga ctgcggctac ccggcctgcc 720ctgagaactg cagcggacac ggcgagtgcg
tgcgcggcgt gtgccagtgc cacgaagact 780tcatgtcgga ggactgcagc gagaagcgct
gtcccggcga ctgcagcggc cacggcttct 840gtgacacggg cgagtgctac tgcgaggagg
gcttcacagg cctggactgt gcccaggtgg 900tcaccccaca gggcctgcag ctgctcaaga
acacggagga ttctctgctg gtgagctggg 960agccctccag ccaggtggat cactacctcc
tcagctacta ccccctgggg aaggagctct 1020ctgggaagca gatccaagtg cccaaggagc
agcacagcta tgagattctt ggtttgctgc 1080ctggaaccaa gtacatagtc accctgcgta
acgtcaagaa tgaagtttct agcagcccac 1140agcatctact tgccaccaca gaccttgctg
tgcttggcac tgcctgggtg acagatgaga 1200ctgagaactc ccttgacgtg gagtgggaaa
acccctcaac tgaggtggac tactacaagc 1260tgcgatatgg ccccatgaca ggacaggagg
tagctgaggt cactgtgccc aagagcagtg 1320accccaagag ccgatatgac atcactggtc
tgcacccggg gactgagtat aagatcacgg 1380tggtgcccat gagaggagag ctggagggca
agccgatcct cctgaatggc aggacagaaa 1440ttgacagtcc aaccaatgtt gtcactgatc
gagtgactga agacacagca actgtctcct 1500gggacccagt gcaggctgtc atagacaagt
atgtagtgcg ctacacttct gctgatgggg 1560acaccaagga aatggcagtg cacaaggatg
agagcagcac tgtcctgacg ggcctgaagc 1620caggagaggc atacaaggtc tacgtgtggg
ctgaaagggg caaccagggg agcaagaaag 1680ctgacaccaa tgccctcaca gaaattgaca
gcccagcaaa cctggtgact gaccgggtga 1740ctgagaatac cgccaccatc tcctgggacc
cggtacaggc caccattgac aagtacgtgg 1800tgcgctacac ctctgctgac gaccaagaga
ccagagaggt tctggtgggg aaggagcaga 1860gcagcactgt cctgacaggc ctgaggccag
gtgtggagta cacagtgcat gtctgggccc 1920agaaggggga ccgagagagc aagaaggctg
acaccaacgc cccgacagat attgacagcc 1980ccaaaaacct ggtgactgac cgggtgacag
agaatatggc cacggtctcc tgggacccgg 2040tgcaggccgc cattgacaag tacgtggtgc
gctacacctc tgctggtgga gagaccaggg 2100aggttccggt ggggaaggag cagagcagca
cagtcctgac aggcctgaga ccgggtatgg 2160agtacatggt gcacgtgtgg gcccagaagg
gggaccagga gagcaagaag gccgacacca 2220aggcccagac agacattgac agcccccaaa
acctggtgac cgaccgggtg acagagaata 2280tggccactgt ctcctgggac ccggtgcggg
ccaccattga caggtatgtg gtgcgctaca 2340cctctgccaa ggacggagag accagggagg
ttccggtggg gaaggagcag agtagcactg 2400tcctgacggg cctgaggccg ggtgtggagt
acacggtgca cgtgtgggcc cagaaggggg 2460cccaggagag caagaaggct gacaccaagg
cccagacaga cattgacagc ccccaaaacc 2520tggtcactga ctgggtgaca gagaatacag
ccactgtctc ctgggacccg gtgcaggcca 2580ccattgacag gtatgtggtg cactacacgt
ctgccaacgg agagaccagg gaggttccag 2640tggggaagga gcagagcagc actgtcctga
cgggcctgag gccgggcatg gagtacacgg 2700tgcacgtgtg ggcccagaag gggaaccagg
agagcaagaa ggctgacacc aaggcccaga 2760cagaaattga cggccccaaa aacctagtga
ctgactgggt gacggagaat atggccactg 2820tctcctggga cccggttcag gccaccattg
acaagtacat ggtgcgctac acctctgctg 2880acggagagac cagggaggtt ccggtgggga
aggagcacag cagcactgtc ctgacgggcc 2940tgagaccagg catggagtac atggtgcacg
tgtgggccca gaagggggcc caggagagca 3000agaaggctga caccaaggcc cagacagaac
tcgaccctcc cagaaacctt cgtccatctg 3060ctgtaacgca gtctggtggc atattgacct
ggacgccccc ctctgctcag atccacggct 3120acattctgac ttaccagttc ccagatggca
cagttaagga gatgcagctg ggacgggaag 3180accagaggtt tgcgttgcaa ggccttgagc
aaggcgccac ctaccctgtc tcccttgttg 3240cctttaaggg tggtcgccgg agcagaaatg
tatccaccac cctctccaca gttggtgccc 3300gtttcccaca cccttcggac tgcagtcagg
ttcagcagaa cagcaatgcc gccagtggtc 3360tgtacaccat ctacctgcat ggcgatgcca
gccggcccct gcaggtgtac tgtgacatgg 3420aaacggacgg aggtggctgg attgtcttcc
agaggcggaa cactgggcag ctggatttct 3480tcaagcgatg gaggagctat gtggaaggct
ttggggaccc catgaaggag ttctggcttg 3540gacttgacaa gctacacaac ctcaccaccg
gcactccagc gcggtatgag gtgagagtgg 3600atttacagac tgccaatgaa tctgcctatg
ctatatatga tttcttccaa gtggcctcca 3660gcaaggagcg gtataagctg acagttggga
aatacagagg cacggcaggg gatgctctta 3720cttaccacaa tggatggaag tttacaactt
ttgacagaga caatgatatc gcactcagca 3780actgtgccct gacacatcat ggtggctggt
ggtataagaa ctgccacttg gccaacccta 3840atggcagata tggggagacc aagcacagtg
agggggtgaa ctgggagcct tggaaaggac 3900atgaattctc cattccttac gtggagttga
aaatccgccc tcatggctac agcagggagc 3960ctgtcctggg cagaaagaag cggacgctga
gaggaaggct gcgaacgttc tgatggcccg 4020tgtgagcagt cctcgcagga gacaccacca
gctgtggcag cttggggcgg ggtgggtagt 4080ggtcactgcg gtctgggagt gctcagatag
cccgcagaac aaatcatgtc accaagcttc 4140aagccatgga ggttccttcc ctctcacctg
catttttgcc cgtctttatg agggtcttga 4200aaatcaaaat agtagttgca cagtatgtgt
aggaaagaca gtactggaac ggcaaggttt 4260ctcagcttat cttcagcaac atatatactg
gattagggca agagaaggaa tcacccagca 4320cttcaccagt tggaaatctc tggaaattta
catctatgta tttaaagttc tgctaatgca 4380aatcttttct ctggaaagaa gcacagagga
ggagttctga tgacccaggg gttagggctg 4440agacaaccgg acgtttgtca cctcctttcc
cattgggttt ttaggaaaac agtgtgaacc 4500tccccctttt aatttctggt gttatgagga
agaataaagg ggataaaagg ggctaagatg 4560gactcatgtt tagctaagtt ctgacttgta
tccagcatgc tggagaccaa agctgccgcc 4620ttactgctat ttttaagtgc cctcttttca
gtcatttgca taattgcgtc catagagctg 4680catatgttgt gaataaattc tcactcattt
caactttgaa taatttgact gtcttgataa 4740ttggttcctc ccaaagactc ttctgcaact
cccattcatg cccaccaggc ctcagactcc 4800ctcttttccc cgccctgcac tattggagcc
ctgggttttg tgggagtgct cagcaccgtg 4860agtcttactg tttgatcgga cagttagcaa
gatcagatcc tttttgctta ttttctatca 4920ctttggaggg ttttctgtag caaaatcagt
gaccaatgaa gtaacttaaa ttcctattga 4980agaaaaaaaa taataaacca cttgattt
500861299PRTHomo sapiens 6Met Ser Leu
Gln Glu Met Phe Arg Phe Pro Met Gly Leu Leu Leu Gly1 5
10 15Ser Val Leu Leu Val Ala Ser Ala Pro
Ala Thr Leu Glu Pro Pro Gly 20 25
30Cys Ser Asn Lys Glu Gln Gln Val Thr Val Ser His Thr Tyr Lys Ile
35 40 45Asp Val Pro Lys Ser Ala Leu
Val Gln Val Asp Ala Asp Pro Gln Pro 50 55
60Leu Ser Asp Asp Gly Ala Ser Leu Leu Ala Leu Gly Glu Ala Arg Glu65
70 75 80Glu Gln Asn Ile
Ile Phe Arg His Asn Ile Arg Leu Gln Thr Pro Gln 85
90 95Lys Asp Cys Glu Leu Ala Gly Ser Val Gln
Asp Leu Leu Ala Arg Val 100 105
110Lys Lys Leu Glu Glu Glu Met Val Glu Met Lys Glu Gln Cys Ser Ala
115 120 125Gln Arg Cys Cys Gln Gly Val
Thr Asp Leu Ser Arg His Cys Ser Gly 130 135
140His Gly Thr Phe Ser Leu Glu Thr Cys Ser Cys His Cys Glu Glu
Gly145 150 155 160Arg Glu
Gly Pro Ala Cys Glu Arg Leu Ala Cys Pro Gly Ala Cys Ser
165 170 175Gly His Gly Arg Cys Val Asp
Gly Arg Cys Leu Cys His Glu Pro Tyr 180 185
190Val Gly Ala Asp Cys Gly Tyr Pro Ala Cys Pro Glu Asn Cys
Ser Gly 195 200 205His Gly Glu Cys
Val Arg Gly Val Cys Gln Cys His Glu Asp Phe Met 210
215 220Ser Glu Asp Cys Ser Glu Lys Arg Cys Pro Gly Asp
Cys Ser Gly His225 230 235
240Gly Phe Cys Asp Thr Gly Glu Cys Tyr Cys Glu Glu Gly Phe Thr Gly
245 250 255Leu Asp Cys Ala Gln
Val Val Thr Pro Gln Gly Leu Gln Leu Leu Lys 260
265 270Asn Thr Glu Asp Ser Leu Leu Val Ser Trp Glu Pro
Ser Ser Gln Val 275 280 285Asp His
Tyr Leu Leu Ser Tyr Tyr Pro Leu Gly Lys Glu Leu Ser Gly 290
295 300Lys Gln Ile Gln Val Pro Lys Glu Gln His Ser
Tyr Glu Ile Leu Gly305 310 315
320Leu Leu Pro Gly Thr Lys Tyr Ile Val Thr Leu Arg Asn Val Lys Asn
325 330 335Glu Val Ser Ser
Ser Pro Gln His Leu Leu Ala Thr Thr Asp Leu Ala 340
345 350Val Leu Gly Thr Ala Trp Val Thr Asp Glu Thr
Glu Asn Ser Leu Asp 355 360 365Val
Glu Trp Glu Asn Pro Ser Thr Glu Val Asp Tyr Tyr Lys Leu Arg 370
375 380Tyr Gly Pro Met Thr Gly Gln Glu Val Ala
Glu Val Thr Val Pro Lys385 390 395
400Ser Ser Asp Pro Lys Ser Arg Tyr Asp Ile Thr Gly Leu His Pro
Gly 405 410 415Thr Glu Tyr
Lys Ile Thr Val Val Pro Met Arg Gly Glu Leu Glu Gly 420
425 430Lys Pro Ile Leu Leu Asn Gly Arg Thr Glu
Ile Asp Ser Pro Thr Asn 435 440
445Val Val Thr Asp Arg Val Thr Glu Asp Thr Ala Thr Val Ser Trp Asp 450
455 460Pro Val Gln Ala Val Ile Asp Lys
Tyr Val Val Arg Tyr Thr Ser Ala465 470
475 480Asp Gly Asp Thr Lys Glu Met Ala Val His Lys Asp
Glu Ser Ser Thr 485 490
495Val Leu Thr Gly Leu Lys Pro Gly Glu Ala Tyr Lys Val Tyr Val Trp
500 505 510Ala Glu Arg Gly Asn Gln
Gly Ser Lys Lys Ala Asp Thr Asn Ala Leu 515 520
525Thr Glu Ile Asp Ser Pro Ala Asn Leu Val Thr Asp Arg Val
Thr Glu 530 535 540Asn Thr Ala Thr Ile
Ser Trp Asp Pro Val Gln Ala Thr Ile Asp Lys545 550
555 560Tyr Val Val Arg Tyr Thr Ser Ala Asp Asp
Gln Glu Thr Arg Glu Val 565 570
575Leu Val Gly Lys Glu Gln Ser Ser Thr Val Leu Thr Gly Leu Arg Pro
580 585 590Gly Val Glu Tyr Thr
Val His Val Trp Ala Gln Lys Gly Asp Arg Glu 595
600 605Ser Lys Lys Ala Asp Thr Asn Ala Pro Thr Asp Ile
Asp Ser Pro Lys 610 615 620Asn Leu Val
Thr Asp Arg Val Thr Glu Asn Met Ala Thr Val Ser Trp625
630 635 640Asp Pro Val Gln Ala Ala Ile
Asp Lys Tyr Val Val Arg Tyr Thr Ser 645
650 655Ala Gly Gly Glu Thr Arg Glu Val Pro Val Gly Lys
Glu Gln Ser Ser 660 665 670Thr
Val Leu Thr Gly Leu Arg Pro Gly Met Glu Tyr Met Val His Val 675
680 685Trp Ala Gln Lys Gly Asp Gln Glu Ser
Lys Lys Ala Asp Thr Lys Ala 690 695
700Gln Thr Asp Ile Asp Ser Pro Gln Asn Leu Val Thr Asp Arg Val Thr705
710 715 720Glu Asn Met Ala
Thr Val Ser Trp Asp Pro Val Arg Ala Thr Ile Asp 725
730 735Arg Tyr Val Val Arg Tyr Thr Ser Ala Lys
Asp Gly Glu Thr Arg Glu 740 745
750Val Pro Val Gly Lys Glu Gln Ser Ser Thr Val Leu Thr Gly Leu Arg
755 760 765Pro Gly Val Glu Tyr Thr Val
His Val Trp Ala Gln Lys Gly Ala Gln 770 775
780Glu Ser Lys Lys Ala Asp Thr Lys Ala Gln Thr Asp Ile Asp Ser
Pro785 790 795 800Gln Asn
Leu Val Thr Asp Trp Val Thr Glu Asn Thr Ala Thr Val Ser
805 810 815Trp Asp Pro Val Gln Ala Thr
Ile Asp Arg Tyr Val Val His Tyr Thr 820 825
830Ser Ala Asn Gly Glu Thr Arg Glu Val Pro Val Gly Lys Glu
Gln Ser 835 840 845Ser Thr Val Leu
Thr Gly Leu Arg Pro Gly Met Glu Tyr Thr Val His 850
855 860Val Trp Ala Gln Lys Gly Asn Gln Glu Ser Lys Lys
Ala Asp Thr Lys865 870 875
880Ala Gln Thr Glu Ile Asp Gly Pro Lys Asn Leu Val Thr Asp Trp Val
885 890 895Thr Glu Asn Met Ala
Thr Val Ser Trp Asp Pro Val Gln Ala Thr Ile 900
905 910Asp Lys Tyr Met Val Arg Tyr Thr Ser Ala Asp Gly
Glu Thr Arg Glu 915 920 925Val Pro
Val Gly Lys Glu His Ser Ser Thr Val Leu Thr Gly Leu Arg 930
935 940Pro Gly Met Glu Tyr Met Val His Val Trp Ala
Gln Lys Gly Ala Gln945 950 955
960Glu Ser Lys Lys Ala Asp Thr Lys Ala Gln Thr Glu Leu Asp Pro Pro
965 970 975Arg Asn Leu Arg
Pro Ser Ala Val Thr Gln Ser Gly Gly Ile Leu Thr 980
985 990Trp Thr Pro Pro Ser Ala Gln Ile His Gly Tyr
Ile Leu Thr Tyr Gln 995 1000
1005Phe Pro Asp Gly Thr Val Lys Glu Met Gln Leu Gly Arg Glu Asp
1010 1015 1020Gln Arg Phe Ala Leu Gln
Gly Leu Glu Gln Gly Ala Thr Tyr Pro 1025 1030
1035Val Ser Leu Val Ala Phe Lys Gly Gly Arg Arg Ser Arg Asn
Val 1040 1045 1050Ser Thr Thr Leu Ser
Thr Val Gly Ala Arg Phe Pro His Pro Ser 1055 1060
1065Asp Cys Ser Gln Val Gln Gln Asn Ser Asn Ala Ala Ser
Gly Leu 1070 1075 1080Tyr Thr Ile Tyr
Leu His Gly Asp Ala Ser Arg Pro Leu Gln Val 1085
1090 1095Tyr Cys Asp Met Glu Thr Asp Gly Gly Gly Trp
Ile Val Phe Gln 1100 1105 1110Arg Arg
Asn Thr Gly Gln Leu Asp Phe Phe Lys Arg Trp Arg Ser 1115
1120 1125Tyr Val Glu Gly Phe Gly Asp Pro Met Lys
Glu Phe Trp Leu Gly 1130 1135 1140Leu
Asp Lys Leu His Asn Leu Thr Thr Gly Thr Pro Ala Arg Tyr 1145
1150 1155Glu Val Arg Val Asp Leu Gln Thr Ala
Asn Glu Ser Ala Tyr Ala 1160 1165
1170Ile Tyr Asp Phe Phe Gln Val Ala Ser Ser Lys Glu Arg Tyr Lys
1175 1180 1185Leu Thr Val Gly Lys Tyr
Arg Gly Thr Ala Gly Asp Ala Leu Thr 1190 1195
1200Tyr His Asn Gly Trp Lys Phe Thr Thr Phe Asp Arg Asp Asn
Asp 1205 1210 1215Ile Ala Leu Ser Asn
Cys Ala Leu Thr His His Gly Gly Trp Trp 1220 1225
1230Tyr Lys Asn Cys His Leu Ala Asn Pro Asn Gly Arg Tyr
Gly Glu 1235 1240 1245Thr Lys His Ser
Glu Gly Val Asn Trp Glu Pro Trp Lys Gly His 1250
1255 1260Glu Phe Ser Ile Pro Tyr Val Glu Leu Lys Ile
Arg Pro His Gly 1265 1270 1275Tyr Ser
Arg Glu Pro Val Leu Gly Arg Lys Lys Arg Thr Leu Arg 1280
1285 1290Gly Arg Leu Arg Thr Phe
129572588DNAHomo sapiens 7gatgggattg gggttttccc ctcccatgtg ctcaagactg
gcgctaaaag ttttgagctt 60ctcaaaagtc tagagccacc gtccagggag caggtagctg
ctgggctccg gggacacttt 120gcgttcgggc tgggagcgtg ctttccacga cggtgacacg
cttccctgga ttggccagac 180tgccttccgg gtcactgcca tggaggagcc gcagtcagat
cctagcgtcg agccccctct 240gagtcaggaa acattttcag acctatggaa actacttcct
gaaaacaacg ttctgtcccc 300cttgccgtcc caagcaatgg atgatttgat gctgtccccg
gacgatattg aacaatggtt 360cactgaagac ccaggtccag atgaagctcc cagaatgcca
gaggctgctc cccccgtggc 420ccctgcacca gcagctccta caccggcggc ccctgcacca
gccccctcct ggcccctgtc 480atcttctgtc ccttcccaga aaacctacca gggcagctac
ggtttccgtc tgggcttctt 540gcattctggg acagccaagt ctgtgacttg cacgtactcc
cctgccctca acaagatgtt 600ttgccaactg gccaagacct gccctgtgca gctgtgggtt
gattccacac ccccgcccgg 660cacccgcgtc cgcgccatgg ccatctacaa gcagtcacag
cacatgacgg aggttgtgag 720gcgctgcccc caccatgagc gctgctcaga tagcgatggt
ctggcccctc ctcagcatct 780tatccgagtg gaaggaaatt tgcgtgtgga gtatttggat
gacagaaaca cttttcgaca 840tagtgtggtg gtgccctatg agccgcctga ggttggctct
gactgtacca ccatccacta 900caactacatg tgtaacagtt cctgcatggg cggcatgaac
cggaggccca tcctcaccat 960catcacactg gaagactcca gtggtaatct actgggacgg
aacagctttg aggtgcgtgt 1020ttgtgcctgt cctgggagag accggcgcac agaggaagag
aatctccgca agaaagggga 1080gcctcaccac gagctgcccc cagggagcac taagcgagca
ctgcccaaca acaccagctc 1140ctctccccag ccaaagaaga aaccactgga tggagaatat
ttcacccttc agatccgtgg 1200gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc
ttggaactca aggatgccca 1260ggctgggaag gagccagggg ggagcagggc tcactccagc
cacctgaagt ccaaaaaggg 1320tcagtctacc tcccgccata aaaaactcat gttcaagaca
gaagggcctg actcagactg 1380acattctcca cttcttgttc cccactgaca gcctcccacc
cccatctctc cctcccctgc 1440cattttgggt tttgggtctt tgaacccttg cttgcaatag
gtgtgcgtca gaagcaccca 1500ggacttccat ttgctttgtc ccggggctcc actgaacaag
ttggcctgca ctggtgtttt 1560gttgtgggga ggaggatggg gagtaggaca taccagctta
gattttaagg tttttactgt 1620gagggatgtt tgggagatgt aagaaatgtt cttgcagtta
agggttagtt tacaatcagc 1680cacattctag gtaggggccc acttcaccgt actaaccagg
gaagctgtcc ctcactgttg 1740aattttctct aacttcaagg cccatatctg tgaaatgctg
gcatttgcac ctacctcaca 1800gagtgcattg tgagggttaa tgaaataatg tacatctggc
cttgaaacca ccttttatta 1860catggggtct agaacttgac ccccttgagg gtgcttgttc
cctctccctg ttggtcggtg 1920ggttggtagt ttctacagtt gggcagctgg ttaggtagag
ggagttgtca agtctctgct 1980ggcccagcca aaccctgtct gacaacctct tggtgaacct
tagtacctaa aaggaaatct 2040caccccatcc cacaccctgg aggatttcat ctcttgtata
tgatgatctg gatccaccaa 2100gacttgtttt atgctcaggg tcaatttctt ttttcttttt
tttttttttt tttctttttc 2160tttgagactg ggtctcgctt tgttgcccag gctggagtgg
agtggcgtga tcttggctta 2220ctgcagcctt tgcctccccg gctcgagcag tcctgcctca
gcctccggag tagctgggac 2280cacaggttca tgccaccatg gccagccaac ttttgcatgt
tttgtagaga tggggtctca 2340cagtgttgcc caggctggtc tcaaactcct gggctcaggc
gatccacctg tctcagcctc 2400ccagagtgct gggattacaa ttgtgagcca ccacgtccag
ctggaagggt caacatcttt 2460tacattctgc aagcacatct gcattttcac cccacccttc
ccctccttct ccctttttat 2520atcccatttt tatatcgatc tcttatttta caataaaact
ttgctgccac ctgtgtgtct 2580gaggggtg
25888393PRTHomo sapiens 8Met Glu Glu Pro Gln Ser
Asp Pro Ser Val Glu Pro Pro Leu Ser Gln1 5
10 15Glu Thr Phe Ser Asp Leu Trp Lys Leu Leu Pro Glu
Asn Asn Val Leu 20 25 30Ser
Pro Leu Pro Ser Gln Ala Met Asp Asp Leu Met Leu Ser Pro Asp 35
40 45Asp Ile Glu Gln Trp Phe Thr Glu Asp
Pro Gly Pro Asp Glu Ala Pro 50 55
60Arg Met Pro Glu Ala Ala Pro Pro Val Ala Pro Ala Pro Ala Ala Pro65
70 75 80Thr Pro Ala Ala Pro
Ala Pro Ala Pro Ser Trp Pro Leu Ser Ser Ser 85
90 95Val Pro Ser Gln Lys Thr Tyr Gln Gly Ser Tyr
Gly Phe Arg Leu Gly 100 105
110Phe Leu His Ser Gly Thr Ala Lys Ser Val Thr Cys Thr Tyr Ser Pro
115 120 125Ala Leu Asn Lys Met Phe Cys
Gln Leu Ala Lys Thr Cys Pro Val Gln 130 135
140Leu Trp Val Asp Ser Thr Pro Pro Pro Gly Thr Arg Val Arg Ala
Met145 150 155 160Ala Ile
Tyr Lys Gln Ser Gln His Met Thr Glu Val Val Arg Arg Cys
165 170 175Pro His His Glu Arg Cys Ser
Asp Ser Asp Gly Leu Ala Pro Pro Gln 180 185
190His Leu Ile Arg Val Glu Gly Asn Leu Arg Val Glu Tyr Leu
Asp Asp 195 200 205Arg Asn Thr Phe
Arg His Ser Val Val Val Pro Tyr Glu Pro Pro Glu 210
215 220Val Gly Ser Asp Cys Thr Thr Ile His Tyr Asn Tyr
Met Cys Asn Ser225 230 235
240Ser Cys Met Gly Gly Met Asn Arg Arg Pro Ile Leu Thr Ile Ile Thr
245 250 255Leu Glu Asp Ser Ser
Gly Asn Leu Leu Gly Arg Asn Ser Phe Glu Val 260
265 270Arg Val Cys Ala Cys Pro Gly Arg Asp Arg Arg Thr
Glu Glu Glu Asn 275 280 285Leu Arg
Lys Lys Gly Glu Pro His His Glu Leu Pro Pro Gly Ser Thr 290
295 300Lys Arg Ala Leu Pro Asn Asn Thr Ser Ser Ser
Pro Gln Pro Lys Lys305 310 315
320Lys Pro Leu Asp Gly Glu Tyr Phe Thr Leu Gln Ile Arg Gly Arg Glu
325 330 335Arg Phe Glu Met
Phe Arg Glu Leu Asn Glu Ala Leu Glu Leu Lys Asp 340
345 350Ala Gln Ala Gly Lys Glu Pro Gly Gly Ser Arg
Ala His Ser Ser His 355 360 365Leu
Lys Ser Lys Lys Gly Gln Ser Thr Ser Arg His Lys Lys Leu Met 370
375 380Phe Lys Thr Glu Gly Pro Asp Ser Asp385
39099205DNAHomo sapiens 9gagaacggcg cgcgccccgc tcggcagctg
cggcggcggc tggtgtaggc gctcacgtca 60gggctgggag gagggggttc gcagctcaga
gagagagaga catgaggcaa cctggcagga 120agcgagaaag agttcggcga agaaggagtg
agttctagag acgccccgcg agcaggaccc 180gcgcctgcag gagagcctgg ccgacccggc
tcctcgcctt ctctgcgcgc tctcccgcgt 240ccgctttcag caccccgagc ggagaacagt
tcccggcagc ccgcagcgct gccgagtggc 300cgccggccgg ccgagtaccc ggagctccag
ggggctcagg agcaccctct gagaacccgc 360tgtgcaccca ccttttcccc tcttttggtg
ggcaagtaag cacgggggaa aagcatccgg 420tggcctcagg gagccctgaa gaaaccgaag
cagaatgtac cagggacaca tgcagaaaag 480taaagaacaa ggatatggaa aactaagcag
tgatgaagac ctcgaaataa ttgttgatca 540aaagcaggga aaaggctcta gggcggcaga
taaggctgtt gccatggtga tgaaggagat 600accgagggag gagtctgctg aagaaaagcc
cctccttact atgacatcac agctggtgaa 660tgagcaacaa gaaagcagac ccctcctgag
tccctccatc gatgactttc tctgtgaaac 720caaatcggaa gcaatagcaa ggccagtaac
atccaataca gctgtattga ccactggctt 780agatctcctc gacctgagtg aaccagtctc
tcaaacccaa accaaagcca agaagtcaga 840gccctcatca aaaacctcat ccctcaagaa
aaaggccgat ggatctgacc tcatcagcac 900ggatgctgag cagagaggcc agcctctcag
agtcccggag acttcatcct tagatctaga 960cattcaaaca caactggaaa aatgggacga
tgttaagttt catggagatc gaaataccaa 1020gggacatcca atggcagaga gaaaatcatc
ctcatctaga actggatcaa aagagctctt 1080atggtcctca gaacacagat ctcaaccaga
actgagtggt ggaaaaagcg ccctcaactc 1140tgagtcggct tcagaattgg aattagtggc
tccgactcag gctcgactga ccaaagaaca 1200tcgctgggga agcgcattac tttctagaaa
ccactcctta gaagaagagt ttgaaagggc 1260aaaagcagca gtggagtcag atacagagtt
ttgggataag atgcaagcag aatgggaaga 1320aatggctcgg aggaactgga tatctgagaa
ccaagaagcc cagaaccaag taaccatctc 1380ggctagtgag aagggatatt actttcacac
tgaaaacccc ttcaaggact ggcctggagc 1440atttgaagaa ggcttaaaaa ggctgaagga
aggggatctg ccagtcacca tcctgttcat 1500ggaagcagca attcttcagg accctggaga
tgcagaggca tggcagttcc tcgggataac 1560ccaggcggag aatgaaaatg aacaagcagc
tattgtcgcc ctccagaggt gcttagaatt 1620acagcccaac aacttaaaag ctttgatggc
cttggctgtg agttatacta acactggcca 1680tcagcaggat gcctgtgacg ctctgaagaa
ttggattaag caaaatccaa agtacaaata 1740ccttgtgaaa agcaagaagg gatctccagg
cctcacccgg cggatgtcta agtccccagt 1800tgatagctct gttctggaag gggtgaagga
attatatctg gaagctgccc accaaaatgg 1860agatatgatc gacccagacc tgcagacagg
tctaggggtt ctgttccacc tgagtggaga 1920atttaataga gcaatagatg catttaacgc
tgccttaact gttcggccag aggactattc 1980actatggaac cgcctcgggg cgaccttggc
gaacggagac cgcagcgagg aagccgtgga 2040ggcctatacg cgagcactgg agattcagcc
aggattcatc cggtccagat acaacctagg 2100aataagctgc atcaacctgg gcgcctacag
agaagcggtc agcaattttc tcactgccct 2160cagtttgcaa agaaagagca ggaatcagca
gcaagttcct catcctgcaa tctctgggaa 2220tatctgggct gccctcagaa ttgcgctctc
tctgatggac caaccagaac tcttccaggc 2280ggctaatctt ggtgacctgg atgtcctctt
aagagctttc aacttggatc cttgaagaaa 2340gaataatacc agtactaata atccctgatc
tgtgtgattg tactgaaaaa tcaaaaacta 2400ttttattatg aatttcaaaa ggataaatca
aatattcaaa aggccatggt catatagccc 2460aaggaaatta attcctgtgg acaatgccca
gtctctgttc agatccaaaa gcacaaaatg 2520ttgtatatag agtcaaagtc aggctcaaaa
gaagaattaa gagactcaag acaaaccaag 2580ataaagtaac tgtgtgttga atactctttc
cacaagttgc aagcatattg caacacatgt 2640tctttgggtt ttttgtctcc ccttgcagct
gatgacacat ctgaggaact tgttcatggg 2700aaacatggaa aaagcactgc ctcagattgg
gaaattctga ctgtttctga actgtcttct 2760tttgcaagac tgaacatagt ttgggccatt
ggtgcatgca catatattaa cttgtgacta 2820aagacagaca ttgcttaaac ctgttccaat
tttaactttt actgtagccc tttgattcca 2880gagagggagc tttgctggca aaagcagttt
ttgcactaga aatttttgct gttcccaatc 2940taaacttttc tggtattgta tatatgcact
ttaatatctt atttatgcct tgctggagtt 3000ttgttttgtt gctccaattt ttaacttggg
ggtgaaagat ttaagaactc agcctcacta 3060gatcaacgga ccaaataaac aattcctgaa
gacagttttt catagtgtag gattttgaag 3120aagtgttttt cttaaagcag atacgatgac
ataaggcgta aatatggcaa acagatcttt 3180tcattcacgt gctttctggg tctgtatgaa
cataagtaag acacaaggta ctgtctcttc 3240atgcattctc tgcaggaaga aagtttgatg
gtatcgtgat ttctattgag ttcaaattaa 3300cttgcttaag tgtcatactt tactggcttt
gccctgcaca ccaatattcc ttttaaacca 3360taaatatttt aaataatatt tcttaagagt
gataaataac attttttcct aaaaattatg 3420ttttcttagt ttcataataa tctgcactgt
tcatcccctt gcattctaag tgatagaaga 3480gcaacacttt caaaccaaat ctcactgaat
gccagtgatt tcatatgaaa gatgcatgcc 3540tctttggctc tttgaacatt tagcaagtat
tgggaatttt ctattagcag tggtgtttta 3600attggttgaa ttccacttag ttctccacag
tgttttgtat gtgtttgggg cttggtgatt 3660attttgaaag atacattttt taaagaatta
ggaacatttt ttgcaaaatg ctaatcagaa 3720tataattttc ccaatgtcag gtatagaagt
tatcccattc ttctttacct ggtctccttc 3780cttcgtctag aaccttcccc acagtagcta
atcctaacaa ctttttattt atgttctttg 3840ataataatag ctcatcccgc tcatgattat
agctacccac aaaaaataaa acaacaacaa 3900caaaaaagac tccacagaat tgggatcaaa
acagtctaat tggctgggga ataagcttct 3960actgaggagt aaatgtgcac acgtgcttat
caattgaatg cgaggtacag gaataaaatc 4020ctctcctctg ttacatctga agtccttggg
ttgtgagagg gcctcaagtt aacagaaacc 4080acaagtcctt gaggcttaca tatgttataa
cctattctag tttataaaga caaaaagctg 4140aggccactgt gaggagtgcc aggttttgat
agtcattgtc aaagttgact tgggccaggt 4200ggtagctcca gctggtcagc agaaggcaga
ggcattcagg cactcatcat gggtagcagc 4260gaccacccag tctcagaccg tgcttctcta
cagtctcagc agcaaagtaa agctcaacaa 4320tgctttcaag aatggtattg taagaattgt
agaattcgag aaacccagtc tggtaaggac 4380tcttggggcc agcctccaag ccatggcagg
gcctgcttga atatgcctga ttaagcatcc 4440tttccttccc tgctccttcc tcctgccatc
tctgtacctt gaccagaaag tcggcatgaa 4500tagttcatgg acaaaggatt gaatagcctc
tgaacaaagg attcaaagca aactttgtaa 4560gtgttctgtg atgccattct atactttgag
cagcagctga atctgcagcc tggatttaaa 4620gcctgaaact agtataaatg tcaaataatt
aatttgttgc taaatggtgg caaattttgt 4680ggcccaagtt ttattttgct attggaacac
aaactccaag gagctgtttt agggaagact 4740aaacaatctt ccattcttac cattatttgg
caatttgcag agcatttagt agtactttct 4800ctagtgtatg tagtatctgt cagatctaag
atttttatag gctgtcaaga agtccagaat 4860tacggcccaa gaggaaaaga gttgcatttt
tcaataatat tttttataaa attaagttaa 4920aaccattatt aaaacatgag tgacaggcca
ggtgcggtgg ctcacgcctg taatcctagc 4980actttgggag gcagaggcgg gcagaccacg
aggtcaggag ttcgagacca gcctgaccaa 5040catggtgaaa ccccgtcttt attaagaata
caaaaattag ccgggtgtag tggtatgcac 5100ctgtaatacc agctactcag gaggctgagg
caggagaatc acttgaacca gggaggcgga 5160ggttgcagtg agccgagatt gcaccgctgc
actccagtct gggtgacaga gcgagactcc 5220gtctcaaaaa aaacaaaaaa gtgacagttt
ataggaatag agcaccacca agaaatcagt 5280agcaagactt tagaacagtc tttttgtata
aatcatatac agtaaatgtt ttgtttctta 5340gaagtgcatc tactttattt tttggcagaa
caatttttgg catcttatct tcatggcatg 5400aaattgtcat ttaaattact ctttccgttt
gaaaggggag ggtatcctta tttcccttct 5460gaggtactct gagaaggctt tgagtttcct
ttttcatgta ttaatgtata acagcctaac 5520agctcctaga agagggtatt cttactagaa
tggctacata cgggtgactt cagttacata 5580caggtgactt ccagggactg acttcgcctg
tagattggat gaaaataaag agaaagccag 5640tgattattgc tgagtttcaa aatataacca
tttctagtag gctacttcta catgctgttg 5700taccttcctg gtgaggagga gacagaggga
taggggagag gaggaggaat aaaaatccct 5760ctcactgggc caatcatact gattcattcc
ctttacttcc tccaaaactc actccccatg 5820actcctgaaa aatgggtaga tctcgtccac
caggaaattt taatagaata ctatgcaccg 5880tgctttctca gtcaatctgg attcctacta
gaacagaaaa gtaaggaaac ataactgggt 5940aggaatgtcc acttacgttt cacaaactca
cttttagagt ttcatgccac tttcctcatt 6000cttctcttta agatactatt atattgaacg
tgggacccat gtctaaatac ttgtatagat 6060aaatatttct tgtatattct tcccatacaa
gcaagacttg aatacagttg tagctagtcc 6120cacttttata ataaaagctt ccaaactctt
tatcataaac tttgtctgtg agcttgggaa 6180tacagagaaa gcaagttgat cattctgaga
cccttcctca tttgccctga atcagttccc 6240caagattgga ctgtacttta agctagttgt
tgtagcagtg gtggttgttg ttttatatct 6300tgagatactt ctagaacatt ctagaaacag
ggtattctaa aggcttactt tatatgccat 6360cctttttgaa ccctcttatt ttagaaatta
ttacacgtgc acacttatga tttcaacctt 6420gtaaaataat ttcaaattca tttcgtatcc
acattttact gcagtttccc tatcatcatc 6480tcaatagtta tagaactggt tgaaattaaa
ctgttttgaa ctaagaagta gatatatata 6540tatatatata tatatatttt tttttttttt
tttaaagagt gttcatagat aaactctggc 6600ataaagtttg taaaaaagca attttttaaa
agcaaaacgt ataacctcag gtacaaaaat 6660attgcatgca ttagtattgc aaatttgcct
actcaaatat taaccaaagc atgcaagata 6720acttgactga atttaaattt acacagtgca
tgattaagtc tcataggtta gcctgttgtt 6780ccttgcctct atacaaggga gcatttttat
gaactttgtg ttacagtttt tctatttgct 6840tttcttatcc tatccttgag attgttttca
tgctatcaca ctgaacttta cagatattca 6900cacatctggt ttctacagtg ttaccagtgc
caaactaatc ctctgaagaa tctgggacat 6960ctaaacactt tttcaaaaac cagttgaggc
acaacagatc ccaaacttaa acaaatccaa 7020gtgtctgaat gctactaaac taaatcatgt
tgactgcctc agcacataaa ggtttcgact 7080ttaaattctt aataatgcat ttcaaaattt
ttctaaaagt ttgggaataa attagaaaaa 7140ctgatattct aaataaggcg aggaggttgg
ggtaatgtca ggcacatcaa aatcatcaca 7200caacgaagta gatatggggc cttaattttc
ataaatgtca catagcaagt gtgtggggta 7260tcacaacatt gttctgatac agccctgttt
gtatggtagt tgagctgctt gggctaaatg 7320tcccttgaga aactggactt gatcaacagg
tctaaaaaca ctgatattta gggagtgatg 7380aggtgatttc acggtcactc agaaccaact
ctgaaatctt ctcactgcct tttcccataa 7440cccctatttt atgaggagac cagaaaggga
gagagcaaag ctcagtgaga ccatgtggtc 7500ttacattagt aacagctttg ttactcactc
catatacccc ggtaccctca cacacatacc 7560atgaactcaa agacagaatt ctacacataa
cacacatagc aaagacgatt gtggggaggc 7620attaaacagt taagaggaaa aaaaaaaaca
cagaaaatta aactgtgagg tttaacaacc 7680tctcatgtgg tattttgcct tgaacgaagt
ttccactacc tgcagcttct gcagaaatac 7740caaaataaca cagtctagag acaagtgtcc
caatgtcaga actgttttct caggaccttc 7800tctataagag aaaactaact tgcaatgctg
cagaatgtca gagcaaaacc ctgatgagct 7860ccgtgggata cttaatagaa gcctgcactc
aaaataatat taagatggaa acaacaacaa 7920caattaggca gttctttgct caactataaa
atgtgtatca ttcattgggg gagggggagg 7980ggtcgtgttc tttggatttc ttggaaaggg
aaatggtata acacaaaaag actaagaggt 8040atacacactc agctcggtaa ttcgagcatt
tggcattttc actgagatat atagcatttc 8100tgcatggata ccatacaaga attctgccat
gtgagaaatg tgttcatgat ggactacatt 8160accagtagtt aagggaaata cagcaaactt
aagtttttac caacttcctt tcatgttccc 8220tatatatcat ctaacccaaa ctttcaattt
acattgccca aatttgttta cattggttga 8280aaaaaaactg attaatttat taaacttttt
acaaactgta acaaacattt aactatttag 8340aaaagacatt tctacatatt ttatatatct
atgaatattg catctctaat ctaatctaaa 8400aatgttgttt tgagacaatt cttctgtgat
gcttcctcct catgctcata aatgtttaaa 8460gtcttgggat ttcattttca ggctgaaata
gataacaaga aaaaaattta agtttaaatg 8520tttacattgc gttactaaat taataccaat
tcaaattaag tgacctatag taaattaatg 8580gcaatggggg ggaggggatt ttaaagtgat
tggcatatta tgtgatactg tatcagtgat 8640ttagtgaaat aatatgtatc atagcacaac
tctgttgggt attaaggctt catcttagta 8700tttccctgcg ttgctctttg aaataaaatt
actaccataa aaataacctc ttatcatatt 8760gatgtgttta atttactcag ttggcttcta
atttgcgtaa caagattaag gtagtatttt 8820tgtactatta ttggaagcat gccttccctt
tttcacatta ttaaattgta tttatatttg 8880tgcaatttta aactatgttt tcaaataaac
tttgtctgcg gcttcgaggt cttttcagga 8940atctttcaaa atgggatttg gggatcagaa
ctccttctga tcaaatggaa tccaatttgt 9000actactggct aaaggtcctt ttattaaata
ttgaatatca ctacatatga ttttgcatga 9060gctatctgga aatcaggaat gcatttttgg
atataaaaca aaactttaaa aatcttcctc 9120ctttgttaat tttttaagac taaaatatta
tttaacctga aattgaattt tgtgattctt 9180tttagaataa aaatattaaa ataaa
920510626PRTHomo sapiens 10Met Tyr Gln
Gly His Met Gln Lys Ser Lys Glu Gln Gly Tyr Gly Lys1 5
10 15Leu Ser Ser Asp Glu Asp Leu Glu Ile
Ile Val Asp Gln Lys Gln Gly 20 25
30Lys Gly Ser Arg Ala Ala Asp Lys Ala Val Ala Met Val Met Lys Glu
35 40 45Ile Pro Arg Glu Glu Ser Ala
Glu Glu Lys Pro Leu Leu Thr Met Thr 50 55
60Ser Gln Leu Val Asn Glu Gln Gln Glu Ser Arg Pro Leu Leu Ser Pro65
70 75 80Ser Ile Asp Asp
Phe Leu Cys Glu Thr Lys Ser Glu Ala Ile Ala Arg 85
90 95Pro Val Thr Ser Asn Thr Ala Val Leu Thr
Thr Gly Leu Asp Leu Leu 100 105
110Asp Leu Ser Glu Pro Val Ser Gln Thr Gln Thr Lys Ala Lys Lys Ser
115 120 125Glu Pro Ser Ser Lys Thr Ser
Ser Leu Lys Lys Lys Ala Asp Gly Ser 130 135
140Asp Leu Ile Ser Thr Asp Ala Glu Gln Arg Gly Gln Pro Leu Arg
Val145 150 155 160Pro Glu
Thr Ser Ser Leu Asp Leu Asp Ile Gln Thr Gln Leu Glu Lys
165 170 175Trp Asp Asp Val Lys Phe His
Gly Asp Arg Asn Thr Lys Gly His Pro 180 185
190Met Ala Glu Arg Lys Ser Ser Ser Ser Arg Thr Gly Ser Lys
Glu Leu 195 200 205Leu Trp Ser Ser
Glu His Arg Ser Gln Pro Glu Leu Ser Gly Gly Lys 210
215 220Ser Ala Leu Asn Ser Glu Ser Ala Ser Glu Leu Glu
Leu Val Ala Pro225 230 235
240Thr Gln Ala Arg Leu Thr Lys Glu His Arg Trp Gly Ser Ala Leu Leu
245 250 255Ser Arg Asn His Ser
Leu Glu Glu Glu Phe Glu Arg Ala Lys Ala Ala 260
265 270Val Glu Ser Asp Thr Glu Phe Trp Asp Lys Met Gln
Ala Glu Trp Glu 275 280 285Glu Met
Ala Arg Arg Asn Trp Ile Ser Glu Asn Gln Glu Ala Gln Asn 290
295 300Gln Val Thr Ile Ser Ala Ser Glu Lys Gly Tyr
Tyr Phe His Thr Glu305 310 315
320Asn Pro Phe Lys Asp Trp Pro Gly Ala Phe Glu Glu Gly Leu Lys Arg
325 330 335Leu Lys Glu Gly
Asp Leu Pro Val Thr Ile Leu Phe Met Glu Ala Ala 340
345 350Ile Leu Gln Asp Pro Gly Asp Ala Glu Ala Trp
Gln Phe Leu Gly Ile 355 360 365Thr
Gln Ala Glu Asn Glu Asn Glu Gln Ala Ala Ile Val Ala Leu Gln 370
375 380Arg Cys Leu Glu Leu Gln Pro Asn Asn Leu
Lys Ala Leu Met Ala Leu385 390 395
400Ala Val Ser Tyr Thr Asn Thr Gly His Gln Gln Asp Ala Cys Asp
Ala 405 410 415Leu Lys Asn
Trp Ile Lys Gln Asn Pro Lys Tyr Lys Tyr Leu Val Lys 420
425 430Ser Lys Lys Gly Ser Pro Gly Leu Thr Arg
Arg Met Ser Lys Ser Pro 435 440
445Val Asp Ser Ser Val Leu Glu Gly Val Lys Glu Leu Tyr Leu Glu Ala 450
455 460Ala His Gln Asn Gly Asp Met Ile
Asp Pro Asp Leu Gln Thr Gly Leu465 470
475 480Gly Val Leu Phe His Leu Ser Gly Glu Phe Asn Arg
Ala Ile Asp Ala 485 490
495Phe Asn Ala Ala Leu Thr Val Arg Pro Glu Asp Tyr Ser Leu Trp Asn
500 505 510Arg Leu Gly Ala Thr Leu
Ala Asn Gly Asp Arg Ser Glu Glu Ala Val 515 520
525Glu Ala Tyr Thr Arg Ala Leu Glu Ile Gln Pro Gly Phe Ile
Arg Ser 530 535 540Arg Tyr Asn Leu Gly
Ile Ser Cys Ile Asn Leu Gly Ala Tyr Arg Glu545 550
555 560Ala Val Ser Asn Phe Leu Thr Ala Leu Ser
Leu Gln Arg Lys Ser Arg 565 570
575Asn Gln Gln Gln Val Pro His Pro Ala Ile Ser Gly Asn Ile Trp Ala
580 585 590Ala Leu Arg Ile Ala
Leu Ser Leu Met Asp Gln Pro Glu Leu Phe Gln 595
600 605Ala Ala Asn Leu Gly Asp Leu Asp Val Leu Leu Arg
Ala Phe Asn Leu 610 615 620Asp Pro625
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