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1. WO2002018567 - REGULATION OF HUMAN AMINOTRANSFERASE-LIKE ENZYME

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[ EN ]

CLAIMS

1. An isolated polynucleotide encoding a aminotransferase-like enzyme polypeptide and being selected from the group consisting of:

a) a polynucleotide encoding a aminotransferase-like enzyme polypeptide comprising an amino acid sequence selected form the group consisting of:

amino acid sequences which are at least about 50% identical to
the amino acid sequence shown in SEQ ID NO. 2,
amino acid sequences which are at least about 50% identical to
the amino acid sequence shown in SEQ ED NO. 9,
the amino acid sequence shown in SEQ ID NO. 2, and
the amino acid sequence shown in SEQ ID NO. 9

b) a polynucleotide comprising the sequence of SEQ ED NO. 1 or SEQ ED NO. 8;

c) a polynucleotide which hybridizes under stringent conditions to a polynucleotide specified in (a) and (b);

d) a polynucleotide the sequence of which deviates from the polynucleotide sequences specified in (a) to (c) due to the degeneration of the genetic code; and

e) a polynucleotide which represents a fragment, derivative or allelic variation of a polynucleotide sequence specified in (a to (d).

2. An expression vector containing any polynucleotide of claim 1.

3. A host cell containing the expression vector of claim 2.

4. A substantially purified aminotransferase-like enzyme polypeptide encoded by a polynucleotide of claim 1.

5. A method for producing a aminotransferase-like enzyme polypeptide, wherein the method comprises the following steps:

a) culturing the host cell of claim 3 under conditions suitable for the expression of the aminotransferase-like enzyme polypeptide; and

b) recovering the aminotransferase-like enzyme polypeptide from the host cell culture.

6. A method for detection of a polynucleotide encoding a aminotransferase-like enzyme polypeptide in a biological sample comprising the following steps:

a) hybridizing any polynucleotide of claim 1 to a nucleic acid material of , a biological sample, thereby forming a hybridization complex; and

b) detecting said hybridization complex.

7. The method of claim 6, wherein before hybridization, the nucleic acid material of the biological sample is amplified.

8. A method for the detection of a polynucleotide of claim 1 or a aminotransferase-like enzyme polypeptide of claim 4 comprising the steps of:

contacting a biological sample with a reagent which specifically interacts with the polynucleotide or the aminotransferase-like enzyme polypeptide.

9. A diagnostic kit for conducting the method of any one of claims 6 to 8.

10. A method of screening for agents which decrease the activity of a aminotransferase-like enzyme, comprising the steps of:

contacting a test compound with any aminotransferase-like enzyme polypeptide encoded by any polynucleotide of claiml;

detecting binding of the test compound to the aminotransferase-like enzyme polypeptide, wherein a test compound which binds to the polypeptide is identified as a potential therapeutic agent for decreasing the activity of a aminotransferase-like enzyme.

11. A method of screening for agents which regulate the activity of a aminotransferase-like enzyme, comprising the steps of:

contacting a test compound with a aminotransferase-like enzyme polypeptide encoded by any polynucleotide of claim 1; and

detecting a aminotransferase-like enzyme activity of the polypeptide, wherein a test compound which increases the aminotransferase-like enzyme activity is identified as a potential therapeutic agent for increasing the activity of the aminotransferase-like enzyme, and wherein a test compound which decreases the aminotransferase-like enzyme activity of the polypeptide is identified as a potential therapeutic agent for decreasing the activity of the aminotransferase- like enzyme.

12. A method of screening for agents which decrease the activity of a aminotransferase-like enzyme, comprising the steps of: contacting a test compound with any polynucleotide of claim 1 and detecting binding of the test compound to the polynucleotide, wherein a test compound which binds to the polynucleotide is identified as a potential therapeutic agent for decreasing the activity of aminotransferase-like enzyme.

13. A method of reducing the activity of aminotransferase-like enzyme, comprising the steps of:

contacting a cell with a reagent which specifically binds to any poly- nucleotide of claim 1 or any aminotransferase-like enzyme polypeptide of claim 4, whereby the activity of aminotransferase-like enzyme is reduced.

14. A reagent that modulates the activity of a aminotransferase-like enzyme polypeptide or a polynucleotide wherein said reagent is identified by the method of any of the claim 10 to 12.

15. A pharmaceutical composition, comprising:

the expression vector of claim 2 or the reagent of claim 14 and a pharmaceutically acceptable carrier.

16. Use of the pharmaceutical composition of claim 15 for modulating the activity of a aminotransferase-like enzyme in a disease.

17. Use of claim 16 wherein the disease is cancer.

18. A cDNA encoding a polypeptide comprising the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9.

19. The cDNA of claim 18 which comprises SEQ ED NO. 1 or SEQ ED NO. 8.

20. The cDNA of claim 18 which consists of SEQ ID NO. 1 or SEQ ED NO. 8.

21. An expression vector comprising a polynucleotide which encodes a polypeptide comprising the amino acid sequence shown in SEQ ID NO. 2 or SEQ ED NO. 9.

22. The expression vector of claim 21 wherein the polynucleotide consists of SEQ ED NO. 1 or SEQ ID NO. 8.

23. A host cell comprising an expression vector which encodes a polypeptide comprising the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO.

24. The host cell of claim 23 wherein the polynucleotide consists of SEQ ED NO. 1 or SEQ ED NO. 8.

25. A purified polypeptide comprising the amino acid sequence shown in SEQ ID NO. 2 or SEQ ED NO. 9.

26. The purified polypeptide of claim 25 which consists of the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9.

27. A fusion protein comprising a polypeptide having the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9.

28. A method of producing a polypeptide comprising the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9, comprising the steps of:

culturing a host cell comprising an expression vector which encodes the polypeptide under conditions whereby the polypeptide is expressed; and isolating the polypeptide.

29. The method of claim 28 wherein the expression vector comprises SEQ ID NO. 1 or SEQ ED NO. 8.

30. A method of detecting a coding sequence for a polypeptide comprising the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9, comprising the steps of:

hybridizing a polynucleotide comprising 11 contiguous nucleotides of SEQ

ID NO. 1 or SEQ ID NO. 8 to nucleic acid material of a biological sample, thereby forming a hybridization complex; and

detecting the hybridization complex.

31. The method of claim 30 further comprising the step of amplifying the nucleic acid material before the step of hybridizing.

32. A kit for detecting a coding sequence for a polypeptide comprising the amino acid sequence shown in SEQ ID NO. 2 or SEQ ED NO. 9, comprising:

a polynucleotide comprising 11 contiguous nucleotides of SEQ ED NO. 1 or SEQ ED NO. 8; and the instructions for the method of claim 30.

33. A method of detecting a polypeptide comprising the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9, comprising the steps of:

contacting a biological sample with a reagent that specifically binds to the polypeptide to form a reagent-polypeptide complex; and

detecting the reagent-polypeptide complex.

34. The method of claim 33 wherein the reagent is an antibody.

35. A kit for detecting a polypeptide comprising the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9, comprising:

an antibody which specifically binds to the polypeptide; and

instructions for the method of claim 33.

36. A method of screening for agents which can modulate the activity of a human aminotransferase-like enzyme, comprising the steps of:

contacting a test compound with a polypeptide comprising an amino acid sequence selected from the group consisting of: (1) amino acid sequences which are at least about 50% identical to the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9 and (2) the amino acid sequence shown in SEQ ED NO. 2 or SEQ ED NO. 9; and

detecting binding of the test compound to the polypeptide, wherein a test compound which binds to the polypeptide is identified as a potential agent for regulating activity of the human aminotransferase-like enzyme.

37. The method of claim 36 wherein the step of contacting is in a cell.

38. The method of claim 36 wherein the cell is in vitro.

39. The method of claim 36 wherein the step of contacting is in a cell-free system.

40. The method of claim 36 wherein the polypeptide comprises a detectable label.

41. The method of claim 36 wherein the test compound comprises a detectable label.

42. The method of claim 36 wherein the test compound displaces a labeled ligand which is bound to the polypeptide.

43. The method of claim 36 wherein the polypeptide is bound to a solid support.

44. The method of claim 36 wherein the test compound is bound to a solid support.

45. A method of screening for agents which modulate an activity of a human aminotransferase-like enzyme, comprising the steps of:

contacting a test compound with a polypeptide comprising an amino acid sequence selected from the group consisting of: (1) amino acid sequences which are at least about 50% identical to the amino acid sequences shown in SEQ ED NO. 2 or SEQ ED NO. 9 and (2) the amino acid sequences shown in SEQ ED NO. 2 and SEQ ED NO. 9; and

detecting an activity of the polypeptide, wherein a test compound which increases the activity of the polypeptide is identified as a potential agent for increasing the activity of the human aminotransferase-like enzyme, and wherein a test compound which decreases the activity of the polypeptide is identified as a potential agent for decreasing the activity of the human aminotransferase-like enzyme.

46. The method of claim 45 wherein the step of contacting is in a cell.

47. The method of claim 45 wherein the cell is in vitro.

48. The method of claim 45 wherein the step of contacting is in a cell-free system.

49. A method of screening for agents which modulate an activity of a human aminotransferase-like enzyme, comprising the steps of:

contacting a test compound with a product encoded by a polynucleotide which comprises the nucleotide sequences shown in SEQ ED NO. 1 or SEQ ID NO. 8; and

detecting binding of the test compound to the product, wherein a test compound which binds to the product is identified as a potential agent for regulating the activity of the human aminotransferase-like enzyme.

50. The method of claim 49 wherein the product is a polypeptide.

51. The method of claim 49 wherein the product is RNA.

52. A method of reducing activity of a human aminotransferase-like enzyme, comprising the step of:

contacting a cell with a reagent which specifically binds to a product encoded by a polynucleotide comprising the nucleotide sequences shown in SEQ ED NO. 1 or SEQ ID NO. 8, whereby the activity of a human aminotransferase- like enzyme is reduced.

53. The method of claim 52 wherein the product is a polypeptide.

54. The method of claim 53 wherein the reagent is an antibody.

55. The method of claim 52 wherein the product is RNA.

56. The method of claim 55 wherein the reagent is an antisense oligonucleotide.

57. The method of claim 56 wherein the reagent is a ribozyme.

58. The method of claim 52 wherein the cell is in vitro.

59. The method of claim 52 wherein the cell is in vivo.

60. A pharmaceutical composition, comprising:

a reagent which specifically binds to a polypeptide comprising the amino acid sequence shown in SEQ D NO. 2 or SEQ ED NO. 9; and

a pharmaceutically acceptable carrier.

61. The pharmaceutical composition of claim 60 wherein the reagent is an antibody.

62. A pharmaceutical composition, comprising:

a reagent which specifically binds to a product of a polynucleotide comprising the nucleotide sequence shown in SEQ ID NO. 1 or SEQ ED NO. 8; and

a pharmaceutically acceptable carrier.

63. The pharmaceutical composition of claim 62 wherein the reagent is a ribozyme.

64. The pharmaceutical composition of claim 62 wherein the reagent is an antisense oligonucleotide.

65. The pharmaceutical composition of claim 62 wherein the reagent is an antibody.

66. A pharmaceutical composition, comprising:

an expression vector encoding a polypeptide comprising the amino acid sequence shown in SEQ ID NO. 2 or SEQ ED NO. 9; and

a pharmaceutically acceptable carrier.

67. The pharmaceutical composition of claim 66 wherein the expression vector comprises SEQ ID NO. 1 or SEQ ED NO. 8.

68. A method of treating a aminotransferase-like enzyme disfunction related disease, wherein the disease is cancer, comprising the step of:

administering to a patient in need thereof a therapeutically effective dose of a reagent that modulates a function of a human aminotransferase-like enzyme, whereby symptoms of the aminotransferase-like enzyme disfunction related disease are ameliorated.

69. The method of claim 68 wherein the reagent is identified by the method of claim 36.

70. The method of claim 68 wherein the reagent is identified by the method of claim 45.