Claims
- 1. A method for identifying a compound that modulates a cold receptor, the method comprising:
(i) contacting a cell comprising a CMR1 polypeptide with the compound, wherein the CMR1 polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid comprising a sequence of SEQ ID NO:2 or 4, and further, wherein the CMR1 polypeptide forms a cation channel; and (ii) determining an effect of the compound upon the cell comprising the CMR1 polypeptide, thereby identifying a compound that modulates the cold receptor.
- 2. The method of claim 1, wherein the effect is a change in intracellular calcium.
- 3. The method of claim 1, wherein the CMR1 polypeptide has the amino acid sequence set out in SEQ ID NO:1 or 3.
- 4. The method of claim 1, wherein the host cell is a non-neuronal cell.
- 5. The method of claim 1, wherein the CMR1 is recombinant.
- 6. The method of claim 1, wherein the nucleic acid has the sequence set out in SEQ ID NO:2 or 4.
- 7. The method of claim 1, further comprising a step of stimulating the cell with menthol.
- 8. The method of claim 1, further comprising a step of stimulating the cell with cold.
- 9. The method of claim 1, wherein the identified compound is an agonist.
- 10. The method of claim 1, wherein the identified compound in an enhancer.
- 11. The method of claim 1, wherein the identified compound is an antagonist.
- 12. The method of claim 1, wherein the cell is derived from mammalian cell lines 293 or CHO.
- 13. The method of claim 1, wherein the nucleic acid that encodes the CMR1 polypeptide has a nucleic acid sequence that has at least 85% identity to SEQ ID NO:2 or 4 by sequence alignment.
- 14. The method of claim 1, wherein the CMR1 polypeptide has at least 85% identity to SEQ ID NO:1 or 3 by sequence alignment.
- 15. The method of claim 1, wherein the CMR-1 polypeptide has at least 90% or greater amino acid sequence identity to SEQ ID NO: 1 or 3 by sequence alignment.
- 16. The method of claim 1, wherein the CMR-1 polypeptide has at least 95% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 17. The method of claim 1, wherein the CMR-1 polypeptide has at least 98% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 18. The method of claim 1, wherein the ability of said compound to modulate cold sensation is confirmed by contacting a mammal with said compound and assessing whether said compound modulates cold sensation.
- 19. The method of claim 18, wherein said mammal is a human.
- 20. A method for identifying a compound that modulates pain, the method comprising:
(i) contacting a cell comprising a CMR1 polypeptide with the compound, wherein the CMR1 polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid comprising a sequence of SEQ ID NO:2 or 4, and further, wherein the CMR1 polypeptide forms a cation channel; and (ii) determining an effect of the compound upon the cell comprising the CMR1 polypeptide, thereby identifying a compound that modulates pain.
- 21. The method of claim 20, wherein the effect is a change in intracellular calcium.
- 22. The method of claim 20, wherein the CMR1 polypeptide has the amino acid sequence set out in SEQ ID NO:1 or 3.
- 23. The method of claim 20, wherein the host cell is a non-neuronal cell.
- 24. The method of claim 20, further wherein the CMR1 is recombinant.
- 25. The method of claim 20, wherein the nucleic acid has the sequence set out in SEQ ID NO:2 or 4.
- 26. The method of claim 20, further comprising a step of stimulating the cell with menthol.
- 27. The method of claim 20, further comprising a step of stimulating the cell with cold.
- 28. The method of claim 20, wherein the identified compound is an agonist.
- 29. The method of claim 20, wherein the identified compound in an enhancer.
- 30. The method of claim 20, wherein the identified compound is an antagonist.
- 31. The method of claim 20, wherein the cell is derived from mammalian cell lines 293 or CHO.
- 32. The method of claim 20, wherein the nucleic acid that encodes the CMR1 polypeptide has a nucleic acid sequence that has at least 85% identity to SEQ ID NO:2 or 4 by sequence alignment.
- 33. The method of claim 20, wherein the CMR1 polypeptide has at least 85% identity to SEQ ID NO:1 or 3 by sequence alignment.
- 34. The method of claim 20, wherein the CMR-1 polypeptide has at least 90% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 35. The method of claim 20, wherein the CMR-1 polypeptide has at least 95% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 36. The method of claim 20, wherein the CMR-1 polypeptide has at least 98% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 37. The method of claim 20, wherein the ability of said compound to modulate cold sensation is confirmed by contacting a mammal with said compound and assessing whether said compound modulates cold sensation.
- 38. The method of claim 37, wherein said mammal is a human.
- 39. A method of modulating cold sensation in a subject, the method comprising the step of administering to the subject an effective amount of a compound identified using the method of claim 1.
- 40. The method of claim 39, wherein the cold sensation is a taste sensation associated with cold foods or cooling compounds.
- 41. A method of modulating pain in a subject, the method comprising the step of administering to the subject an effective amount of a compound identified using the method of claim 20.
- 42. A compound capable of modulating a cold receptor, the compound identified using the method of claim 1.
- 43. A compound capable of modulating a pain, the compound identified using the method of claim 20.
- 44. An isolated nucleic acid encoding a CMR1 polypeptide, wherein the polypeptide comprises at least 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 or 3.
- 45. An isolated nucleic acid of claim 44, wherein the polypeptide comprises the sequence set forth in SEQ ID NO:1 or 3.
- 46. An isolated nucleic acid of claim 44, wherein the nucleic acid comprises the sequence set forth in SEQ ID NO:2 or 4.
- 47. An isolated nucleic acid encoding a CMR1 polypeptide, wherein the polypeptide comprises at least 98% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 or 3.
- 48. An isolated CMR1 polypeptide comprising at least 95% identity to the amino acid sequence set forth in SEQ ID NO:1 or 3.
- 49. An isolated CMR1 polypeptide of claim 48, wherein the polypeptide comprises the amino acid sequence set forth in SEQ ID NO:1 or 3.
- 50. An isolated CMR1 polypeptide comprising at least 98% identity to the amino acid sequence set forth in SEQ ID NO:1 or 3.
- 51. An expression vector comprising a nucleic acid sequence encoding a CMR1 polypeptide that has at least 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 or 3.
- 52. A host cell comprising an expression vector of claim 51.
- 53. An expression vector comprising a nucleic acid sequence encoding a CMR1 polypeptide that has at least 98% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 or 3.
- 54. A host cell comprising an expression vector of claim 51.
- 55. A method for identifying a compound that imparts a menthol-like taste, the method comprising the steps of:
(i) contacting a cell comprising a CMR1 polypeptide with the compound, wherein the CMR1 polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid comprising a sequence of SEQ ID NO:2 or 4, and further, wherein the CMR1 polypeptide forms a cation channel; (ii) determining an effect of the compound upon the cell comprising the CMR1 polypeptide; and (iii) performing taste tests with said compound, thereby identifying a compound that imparts a menthol-like taste.
- 56. The method of claim 55, wherein the effect is a change in intracellular calcium.
- 57. The method of claim 55, wherein the CMR1 polypeptide has the amino acid sequence set out in SEQ ID NO:1 or 3.
- 58. The method of claim 55, wherein the host cell is a non-neuronal cell.
- 59. The method of claim 55, wherein the CMR1 is recombinant.
- 60. The method of claim 55, wherein the nucleic acid has the sequence set out in SEQ ID NO:2 or 4.
- 61. The method of claim 55, further comprising a step of stimulating the cell with menthol.
- 62. The method of claim 55, further comprising a step of stimulating the cell with cold.
- 63. The method of claim 55, wherein the identified compound is an agonist.
- 64. The method of claim 55, wherein the identified compound in an enhancer.
- 65. The method of claim 55, wherein the identified compound is an antagonist.
- 66. The method of claim 55, wherein the cell is derived from mammalian cell lines 293 or CHO.
- 67. The method of claim 55, wherein the nucleic acid that encodes the CMR1 polypeptide has a nucleic acid sequence that has at least 85% identity to SEQ ID NO:2 or 4 by sequence alignment.
- 68. The method of claim 55, wherein the CMR1 polypeptide has at least 85% identity to SEQ ID NO:1 or 3 by sequence alignment.
- 69. The method of claim 55, wherein the CMR-1 polypeptide has at least 90% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 70. The method of claim 55, wherein the CMR-1 polypeptide has at least 95% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 71. The method of claim 55, wherein the CMR-1 polypeptide has at least 98% or greater amino acid sequence identity to SEQ ID NO:1 or 3 by sequence alignment.
- 72. The method of claim 55, wherein said mammal is a human.
- 73. A method for identifying a compound that modulates a cold receptor, the method comprising the steps of:
(i) contacting a cell comprising a CMR1 polypeptide with an agonist and the compound, wherein the CMR1 polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid comprising a sequence of SEQ ID NO:2 or 4, and further, wherein the CMR1 polypeptide forms a cation channel; (ii) determining an effect of the compound upon the cell comprising the CMR1 polypeptide; and (iii) performing taste tests with said compound, thereby identifying a compound that modulates the cold receptor.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Ser. No. 60/351,974, filed Jan. 25, 2002, and U.S. Ser. No. 60355,037, filed Feb. 7, 2002, which are herein incorporated by reference in their entirety.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT
[0002] This invention was made with government support under grant number GM44298 awarded by the National Institutes of Health. The Government has certain rights in this invention.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60355037 |
Feb 2002 |
US |
|
60351974 |
Jan 2002 |
US |