Claims
- 1. An isolated EER-7 protein having an amino acid sequence comprising at least 10 contiguous amino acids from the sequence depicted in SEQ ID NO:2 or which has at least 60% sequence similarity with SEQ ID NO:2, which EER-7 protein has (i) lysyl oxidase activity; (ii) comprises four copies of a SRCR domain having a sequence greater than 80% similar to a sequence selected from the group consisting of SEQ ID NOs: 3, 4, 5, and 6; and (iii) comprises a conserved catalytic domain of lysyl oxidase enzymes having a sequence as depicted inSEQ ID NO:7.
- 2. The EER-7 protein of claim 1, wherein the protein has specific binding activity with an anti-EER-7 antibody.
- 3. The EER-7 protein of claim 1 which is a human EER-7 protein.
- 4. The EER-7 protein of claim 3 which has an amino acid sequence as depicted in SEQ ID NO: 2.
- 5. The EER-7 protein of claim 3 which is encoded by a nucleic acid having a sequence as depicted in SEQ ID NO: 1.
- 6. The EER-7 protein of claim 1 having at least 60% sequence identity to human EER-7 protein having an amino acid sequence as depicted in SEQ ID NO: 2.
- 7. A polypeptide fragment of an EER-7 protein, wherein the fragment has a property selected from the group consisting of:
a) comprising from one to four copies of a SRCR domain having a sequence greater than 80% similar to a sequence selected from the group consisting of SEQ ID Nos: 3, 4, 5, and 6; b) a conserved catalytic domains of lysyl oxidase enzymes having a sequence as depicted in SEQ ID NO: 7; c) specific binding activity with an anti-EER-7 antibody; and d) any combination thereof.
- 8. An isolated nucleic acid encoding the EER-7 protein of claim 1.
- 9. The nucleic acid of claim 8 which is a cDNA.
- 10. The nucleic acid of claim 8, wherein the EER-7 protein is a human EER-7 protein.
- 11. The EER-7 protein of claim 10 which has an amino acid sequence as depicted in SEQ ID NO: 2.
- 12. The nucleic acid of claim 8 which comprises a nucleotide sequence as depicted in SEQ ID NO: 1.
- 13. A vector comprising a nucleic acid encoding a fragment of an EER-7 protein operatively associated with an expression control sequence, wherein the fragment has a property selected from the group consisting of:
a) comprising from one to four copies of a SRCR domain having a sequence greater than 80% similar to a sequence selected from the group consisting of SEQ ID Nos: 3, 4, 5, and 6; b) a conserved catalytic domains of lysyl oxidase enzymes having a sequence as depicted in SEQ ID NO: 7; c) specific binding activity with an anti-EER-7 antibody; and d) any combination thereof.
- 14. The vector according to claim 13, wherein the fragment of an EER-7 protein is a full length EER-7 protein.
- 15. A host cell transfected with the vector of claim 14.
- 16. A non-human animal transformed with the vector of claim 14, wherein the animal expresses an EER-7 protein at a detectable level in response to estrogen.
- 17. A method for producing EER-7 protein, which method comprises isolating EER-7 protein produced by the host cells of claim 15, wherein the host cells have been cultured under conditions that provide for expression of the EER-7 protein by the vector.
- 18. An isolated nucleic acid of at least 20 bases that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence as depicted in SEQ ID NO: 1.
- 19. The nucleic acid of claim 18, wherein at least ten nucleotides are contiguous nucleotides from the nucleic acid sequence as depicted in SEQ ID NO: 1.
- 20. The nucleic acid of claim 18 which is detectably labeled.
- 21. An antibody that specifically binds to the EER-7 protein of claim 1.
- 22. A method for detecting an EER-7 protein, which method comprises detecting binding of the antibody of claim 21 to a molecule in a sample suspected of containing an EER-7 protein, wherein the antibody is contacted with the sample under conditions that perrmit specific binding with any EER-7 protein present in the sample and binding of the antibody to the molecule in the sample indicates the presence of EER-7.
- 23. A method for detecting expression of EER-7, which method comprises detecting mRNA encoding EER-7 in a sample from a cell suspected of expressing EER-7.
- 24. The method according to claim 23 wherein mRNA encoding EER-7 is detected by hybridization to an EER-7-specific nucleic acid.
- 25. The method according to claim 24 wherein the EER-7-specific nucleic acid is at least 10 nucleotides in length and has a sequence identical to a sequence of the same number of bases in SEQ ID NO: 1, or the complementary sequence thereof.
- 26. An assay system for identifying selective estrogen receptor ligands, comprising transformed cells that express different functional estrogen receptors, wherein the number of cells is sufficient to transcribe a detectable amount of mRNA encoding EER-7.
- 27. The assay system of claim 26, wherein the estrogen receptor is a human estrogen receptor.
- 28. The assay system of claim 26 which is an endothelial cell.
- 29. The assay system of claim 28 which is a human umbilical vein cell.
- 30. A method for identifying a compound that selectively regulates EER-7 mRNA transcription through an estrogen receptor, which method comprises detecting a difference in the level of EER-7 mRNA in an assay system of claim 26 contacted with a test compound, wherein a difference in the level of EER-7 mRNA indicates that the test compound selectively regulates the estrogen receptor.
- 31. The method according to claim 30, wherein the test compound is an estrogen or an estrogen analog.
- 32. The method according to claim 31, wherein the test compound is an estrogen receptor selective agonist or antagonist.
- 33. The method according to claim 30, wherein the level of mRNA decreases when contacted with a test compound that regulates expression through the estrogen receptor.
- 34. The method according to claim 30, wherein the level of mRNA increases when contacted with a test compound that regulates expression through the estrogen receptor.
- 35. The method according to claim 30, wherein the estrogen receptor is a human estrogen receptor.
- 36. The method according to claim 30, wherein the first estrogen receptor is an ERα.
- 37. The method according to claim 36, wherein the second estrogen receptor is an ERβ.
- 38. The method according to claim 30, wherein the cell is an endothelial cell.
- 39. The method according to claim 38, wherein the cell is a human umbilical vein cell.
- 40. The polypeptide fragment of claim 7, wherein the four copies of SRCR domains comprise the sequences as depicted in SEQ ID NOS: 3-6.
- 41. The polypeptide fragment of claim 7, having at least 46% sequence similarity to the catalytic domain of lysyl oxidase enzyme having an amino acid sequence as depicted as SEQ ID NO: 7.
- 42. The assay system of claim 26, wherein the transformed cells comprise two different populations.
- 43. The assay system of claim 42, wherein one population expresses the ERα estrogen receptor.
- 44. The assay system of claim 43, wherein the other population expresses the ERβ estrogen receptor.
- 45. A non-human EER-7 knockout animal, wherein endogenous EER-7 expression is suppressed in the animal.
- 46. A non-human animal transformed with a vector comprising a nucleic acid encoding a protein that regulates EER-7 expression, wherein the protein is operatively associated with an expression control sequence; wherein the animal expresses an EER-7 protein at a detectable level in response to estrogen.
- 47. A non-human animal transformed with a vector comprising a nucleic acid encoding the EER-7 protein of claim 1, wherein the animal expresses the EER-7 protein at a detectable level in response to estrogen.
- 48. An isolated polypeptide selected from the group consisting of: a) a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO: 2; b) an allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule consisting of SEQ ID NO: 1 under stringent conditions; c) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 1; d) a polypeptide comprising an amino acid sequence which is at least 60% identical to the amino acid sequence of SEQ ID NO: 2.
- 49. A method for identifying a compound which binds to a polypeptide of claim 48 which method comprises determining whether a test compound contacted with the polypeptide binds to the polypeptide.
- 50. The method of claim 49 wherein the polypeptide is not in or on the cell.
- 51. The method of claim 49 wherein the polypeptide is in or on the cell.
- 52. The method of claim 49 wherein the binding of the test compound to the polypeptide is detected by directly detecting binding of a test compound to the polypeptide.
- 53. The method of claim 49 wherein the binding of the test compound to the polypeptide is detected by detecting lysyl oxidase activity, wherein a change in lysyl oxidase activity indicates binding of the test compound to the polypeptide.
- 54. A method for modulating the activity of a polypeptide of claim 48, which method comprises contacting the polypeptide with a compound which binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide.
- 55. The method of claim 54 wherein the polypeptide is not in or on the cell.
- 56. The method of claim 54 wherein the polypeptide is in or on the cell.
- 57. A method for identifying a compound which modulates the activity of a polypeptide of claim 48, which method comprises contacting the polypeptide with a test compound and determining the effect of the test compound on the activity of the polypeptide, wherein a change in the activity of the polypeptide indicates that the compound modulates the activity of the polypeptide.
PRIORITY
[0001] This application is a divisional of U.S. patent application Ser. No. 09/924,946 filed Aug. 8, 2001 which claims priority under 35 U.S.C. § 119 from U.S. Provisional Patent Application Serial No. 60/223,763, filed Aug. 8, 2000 and U.S. Provisional Patent Application Serial No. 60/255,838, filed Dec. 15, 2000; each of which is hereby incorporated by reference in its entirety.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60223763 |
Aug 2000 |
US |
|
60255838 |
Dec 2000 |
US |
Divisions (1)
|
Number |
Date |
Country |
Parent |
09924946 |
Aug 2001 |
US |
Child |
10456982 |
Jun 2003 |
US |