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1. (WO2007002641) METHOD FOR IDENTIFYING CART RECEPTOR AND USES THEREOF
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CLAIMS

1. A method for determining if a test compound interacts with GHSR, comprising contacting (a) said test compound, and (b) (i) CART or (ii) a CART derivative which interacts with GHSR, and comparing interaction of (b) with GHSR in the presence of (a), to its interaction with GHSR in the absence of (a), as a determination of whether said test compound interacts with GHSR.

2. The method of claim 1 , wherein (b) is labeled with a detectable moiety.

3. The method of claim 1, wherein said GHSR is presented on a cell surface.

4. The method of claim 3, comprising determining a downstream activity of GHSR as a determination of said interaction.

5. The method of claim 3, wherein said cell expresses GHSR endogenously.

6. The method of claim 3, wherein said cell has been transformed or transfected with an isolated nucleic acid molecule which encodes GHSR.

7. The method of claim 3, wherein said cell is a eukaryote.

8. The method of claim 3, wherein said cell is a prokaryote.

9. The method of claim 3, comprising contacting said cell with said test compound and CART or GHSR binding fragment of CART simultaneously.

10. The method of claim 3, comprising contacting said cell with said test compound and CART or GHSR binding fragment of CART sequentially.

11. A method for determining if a test compound modulates GHSR activity comprising contacting said compound to a cell which has been transformed or transfected with
(a) a nucleic acid molecule which comprises:
(i) a nucleotide sequence which encodes GHSR,
(ii) a nucleotide sequence encoding a cleavage site for a protease or a portion of a protease, and (iii) a nucleotide sequence wήicn encodes a piυicm wmuu activates a reporter gene in said cell, and
(b) a nucleic acid molecule which comprises:
(i) a nucleotide sequence which encodes a test protein whose interaction with said GHSR in the presence of said test compound is to be measured, and
(ii) a nucleotide sequence which encodes a protease or a portion of a protease which is specific for said cleavage site,
and determining activity of said reporter gene as a determination of whether said compound modulates GHSR activity.

12. The method of claim 11, further comprising contacting said cell with CART, a GHSR binding fragment thereof, ghrelin, or a GHSR binding fragment thereof.

13. The method of claim 11, wherein said protease or portion of a protease is tobacco etch virus nuclear inclusion A protease.

14. The method of claim 11, wherein said protein which activates said reporter gene is a transcription factor.

15. The method of claim 14, wherein said transcription factor is tTA or GAL4.

16. The method of claim 11 , wherein said test protein is an inhibitory protein.

17. The method of claim 16, wherein said inhibitory protein is an arrestin.

18. The method of claim 11 , wherein said cell is a eukaryote.

19. The method of claim 115 wherein said reporter gene is an exogenous gene.

20. The method of claim 19, wherein said exogenous gene encodes β-galactosidase, β- lactamase or luciferase.

21. The method of claim 11, wherein the nucleotide sequence encoding GHSR is modified to increase interaction with said test protein.

22. The method of claim 11, wherein said modification comprises replacing all or part of the nucleotide sequence of the C-terminal region of GHSR with a nucleotide sequence which encodes an ammo acid sequence which has higher altmity tor said second test protein than the original sequence.

23. The method of claim 22, wherein the nucleotide sequence of said C-terminal region is replaced by a nucleotide sequence encoding all or a part of the C- terminal region of AVPR2, AGTRLI5 GRPR, F2RL1, CXCR2/IL-8B, CCR4, or GRPR.

24. The method of claim 11, comprising contacting more than one compound to a plurality of samples of cells, each of said samples being contacted by one or more of said compounds, wherein each of said cell samples have been transformed or transfected with (a) and (b), and determining activity of reporter genes in said plurality of said samples to determine if any of said compounds interacts with GHSR.

25. The method of claim 21, comprising contacting each of said samples with one compound, each of which differs from all others.

26. The method of claim 21, comprising contacting each of said samples with a mixture of said compounds.

27. A method for determining if a test compound is a CART inhibitor, comprising contacting said test compound and CART to GHSR or a GHSR fragment, determining activity of CART in the presence of said test compound to activity of CART in its absence, wherein a decrease in CART activity in the presence of said test compound is indicative of a CART inhibitor.

28. The method of claim 27, wherein said CART activity is binding to GHSR or GHSR fragment.

29. The method of claim 28, wherein said CART is labeled.

30. The method of claim 27, wherein said GHSR or GHSR fragment is expressed by a cell, and said activity of CART is a downstream property following binding of CART thereto.

31. A method for identifying an analogue of CART, comprising contact a test compound to GHSR or a fragment of GHSR, and determining if said test compound binds to said GHSR or fragment of GHSR and exhibits a property exhibited by CART upon binding to GHSR or said fragment of GHSR, wherein presence of said property indicates said test compound is a CART analogue.

32. The method of claim 31 , wherein said test compound exhibits greater activity than CART upon said binding.

33. The method of claim 31, comprising contacting said test compound and CART, to GHSR or said GHSR fragment, wherein one of CART and said test compound are labeled, and determining label bound to GHSR or said GHSR fragment, to binding of said label in the absence of unlabelled substance.

34. The method of claim 31, wherein said GHSR or fragment of GHSR is expressed by a cell.

35. The method of claim 34, comprising determining a downstream property following binding of CART and said test compound thereto.

36. A method for modulating activity of a GHSR, comprising contacting said GHSR with a substance which binds to GHSR but is not ghrelin, in an amount sufficient to bind to GHSR and modulate its activity.

37. The method of claim 36, wherein said substance is a non-orthosteric binding partner for GHSR.

38. The method of claim 37, wherein said substance is an allosteric modifier of GHSR.

39. The method of claim 38, wherein said substance is CART or a CART derivative.

40. The method of claim 39, wherein said CART derivative is CART 55-102.

41. Recombinant cell, transformed or transfected with:
(a) a nucleic acid molecule which comprises:
(i) a nucleotide sequence which encodes GHSR, (ii) a nucleotide sequence encoding a cleavage site for a protease or a portion of a protease, and
(iii) a nucleotide sequence which encodes a protein which activates a reporter gene in said cell, and
(b) a nucleic acid molecule which comprises:
(i) a nucleotide sequence which encodes a test protein whose interaction with GHSR in the presence of said test compound is to be measured, and
(ii) a nucleotide sequence which encodes a protease or a portion of a protease which is specific for said cleavage site.

42. The recombinant cell of claim 41, wherein one or both of said nucleic acid molecules are stably incorporated into the genome of said cell.

43. The recombinant cell of claim 41, wherein said cell has been transformed or transfected with said reporter gene.

44. The recombinant cell of claim 41 , wherein said protease or portion of a protease is tobacco etch virus nuclear inclusion A protease.

45. The recombinant cell of claim 41, wherein said protein which activates said reporter gene is a transcription factor.

46. The recombinant cell of claim 41, wherein said transcription factor is tTA or GAL4.

47. The recombinant cell of claim 41, wherein said second protein is an inhibitory protein.

48. The recombinant cell of claim 41 , wherein said inhibitory protein is an arrestin.

49. The recombinant cell of claim 41, wherein said cell is a eukaryote.

50. The recombinant cell of claim 41 , wherein said cell is a prokaryote.

51. The recombinant cell of claim 41, wherein said reporter gene is an exogenous gene.

52. The recombinant cell of claim 41, wherein said exogenous gene encodes β- galactosidase, β-lactamase or luciferase.

53. The recombinant cell of claim 41, wherein the nucleotide sequence encoding GHSR is modified to increase interaction with said test protein.

54. The recombinant cell of claim 53, wherein said modification comprises replacing all or part of the nucleotide sequence of the C-terminal region of GHSR with a nucleotide sequence which encodes an amino acid sequence which has higher affinity for said test protein than the original sequence.

55. The recombinant cell of claim 54, wherein the nucleotide sequence of said C- terminal region is replaced by a nucleotide sequence encoding the C-terminal region of AVPR2, AGTRLI, GRPR, F2RL1, CXCR2/IL-8B, or CCR4.

56. An isolated nucleic acid molecule which comprises, in 5' to 3 ' order,
(i) a nucleotide sequence which encodes GHSR,
(ii) a nucleotide sequence encoding a cleavage site for a protease or a portion of a protease, and
(iii) a nucleotide sequence which encodes a protein which activates a reporter gene in said cell.

57. The isolated nucleic acid molecule of claim 56, wherein said protease or portion of a protease is tobacco etch virus nuclear inclusion A protease.

58. The isolated nucleic acid molecule of claim 56, wherein said protein which activates said reporter gene is a transcription factor.

59. The isolated nucleic acid molecule of claim 58, wherein said transcription factor is tTA or GAL4.

60. Expression vector comprising the isolated nucleic acid molecule of claim 56, operably linked to a promoter.

61. A fusion protein produced by expression of the isolated nucleic acid molecule of claim 56.

62. A test kit useful for determining if a test compound modulates GHSR activity comprising a separate portion of each of:
(a) a nucleic acid molecule which comprises:
(i) a nucleotide sequence which encodes GHSR,
(ii) a nucleotide sequence encoding a cleavage site for a protease or a portion of a protease,
(iii) a nucleotide sequence which encodes a protein which activates a reporter gene in said cell, and
(b) a nucleic acid molecule which comprises:
(i) a nucleotide sequence which encodes a test protein whose interaction with said GHSR in the presence of said test compound is to be measured,
(ii) a nucleotide sequence which encodes a protease or a portion of a protease which is specific for said cleavage site, and
(c) container means for holding each of (a) and (b) separately from each other.

63. The test kit of claim 62, wherein said protease or portion of a protease is tobacco etch virus nuclear inclusion A protease.

64. The test kit of claim 62, wherein said protein which activates said reporter gene is a transcription factor.

65. The test kit of claim 64, wherein said transcription factor is tTA or GAL4.

66. The test kit of claim 62, wherein said test protein is an inhibitory protein.

67. The test kit of claim 66, wherein said inhibitory protein is an arrestin.

68. The test kit of claim 66, further comprising a separate portion of an isolated nucleic acid molecule which encodes a reporter gene.

69. The test kit of claim 68, wherein said reporter gene encodes β-galactosidase, - lactamase or luciferase.

70. The test kit of claim 62, wherein the nucleotide sequence encoding GHSR is modified to increase interaction with said second test protein.

71. The test kit of claim 70, wherein said modification comprises replacing all or part of the nucleotide sequence of the C-terminal region of GHSR with a nucleotide sequence which encodes an amino acid sequence which has higher affinity for said second test protein than the original sequence.

72. The test kit of claim 71, wherein said nucleotide sequence of said C-terminal region is replaced by a nucleotide sequence encoding the C-terminal region of AVPR2, AGTRLI, GRPR, F2RL1, CXCR2/IL-8B or CCR4.

73. The test kit of claim 62, further comprising a separate portion of ghrelin, a ghrelin derivative which binds to GHSR, CART, or a CART derivative which binds to GHSR.