Claims
- 1. An isolated nucleic acid molecule that encodes a polypeptide having mu3 opiate receptor activity.
- 2. The isolated nucleic acid molecule of claim 1, wherein said isolated nucleic acid molecule comprises a nucleic acid sequence with a length and a percent identity to the sequence set forth in SEQ ID NO: 1 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (81, 100), point B has coordinates (81, 65), point C has coordinates (15, 65), and point D has coordinates (15, 100).
- 3. The isolated nucleic acid molecule of claim 1, wherein said polypeptide comprises an amino acid sequence with a length and a percent identity to the sequence set forth in SEQ ID NO: 2 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (26, 100), point B has coordinates (26, 65), point C has coordinates (5, 65), and point D has coordinates (5, 100).
- 4. The isolated nucleic acid molecule of claim 1, wherein said isolated nucleic acid molecule hybridizes under hybridization conditions to the sense or antisense strand of the sequence set forth in SEQ ID NO: 1 or 3.
- 5. The isolated nucleic acid molecule of claim 1, wherein said isolated nucleic acid molecule comprises the sequence set forth in SEQ ID NO: 4, 6, 8, or 10.
- 6. An isolated nucleic acid molecule that hybridizes under hybridization conditions to the sense or antisense strand of a nucleic acid that encodes a polypeptide having mu3 opiate receptor activity, wherein said isolated nucleic acid molecule is at least 12 nucleotides in length, and wherein said isolated nucleic acid molecule does not hybridize to the sense or antisense strand of the sequence set forth in SEQ ID NO: 12 or 13.
- 7. An isolated nucleic acid molecule comprising a nucleic acid sequence with a length and a percent identity to the sequence set forth in SEQ ID NO: 1 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (81, 100), point B has coordinates (81, 65), point C has coordinates (15, 65), and point D has coordinates (15, 100).
- 8. The isolated nucleic acid molecule of claim 7, wherein said isolated nucleic acid molecule encodes a polypeptide having mu3 opiate receptor activity.
- 9. A cell comprising an isolated nucleic acid molecule that encodes a polypeptide having mu3 opiate receptor activity.
- 10. The cell of claim 9, wherein said isolated nucleic acid molecule comprises a nucleic acid sequence with a length and a percent identity to the sequence set forth in SEQ ID NO: 1 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (81, 100), point B has coordinates (81, 65), point C has coordinates (15, 65), and point D has coordinates (15, 100).
- 11. The cell of claim 9, wherein said polypeptide comprises an amino acid sequence with a length and a percent identity to the sequence set forth in SEQ ID NO: 2 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (26, 100), point B has coordinates (26, 65), point C has coordinates (5, 65), and point D has coordinates (5, 100).
- 12. The cell of claim 9, wherein said isolated nucleic acid molecule hybridizes under hybridization conditions to the sense or antisense strand of the sequence set forth in SEQ ID NO: 1 or 3.
- 13. The cell of claim 9, wherein said isolated nucleic acid molecule comprises the sequence set forth in SEQ ID NO: 4, 6, 8, or 10.
- 14. A cell comprising an isolated nucleic acid molecule that hybridizes under hybridization conditions to the sense or antisense strand of a nucleic acid that encodes a polypeptide having mu3 opiate receptor activity, wherein said isolated nucleic acid molecule is at least 12 nucleotides in length, and wherein said isolated nucleic acid molecule does not hybridize to the sense or antisense strand of the sequence set forth in SEQ ID NO: 12 or 13.
- 15. A substantially pure polypeptide having mu3 opiate receptor activity.
- 16. The substantially pure polypeptide of claim 15, wherein said polypeptide is encoded by a nucleic acid sequence having a length and a percent identity to the sequence set forth in SEQ ID NO: 1 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (81, 100), point B has coordinates (81, 65), point C has coordinates (15, 65), and point D has coordinates (15, 100).
- 17. The substantially pure polypeptide of claim 15, wherein said polypeptide comprises an amino acid sequence with a length and a percent identity to the sequence set forth in SEQ ID NO: 2 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (26, 100), point B has coordinates (26, 65), point C has coordinates (5, 65), and point D has coordinates (5, 100).
- 18. The substantially pure polypeptide of claim 15, wherein said polypeptide is encoded by a nucleic acid molecule that hybridizes under hybridization conditions to the sense or antisense strand of the sequence set forth in SEQ ID NO: 1 or 3.
- 19. The substantially pure polypeptide of claim 15, wherein said polypeptide comprises the sequence set forth in SEQ ID NO: 5, 7, 9, or 11.
- 20. A method for identifying a mu3 opiate receptor agonist, said method comprising:
a) contacting a cell with a test molecule, wherein said cell comprises an isolated nucleic acid molecule that encodes a polypeptide having mu3 opiate receptor activity, and wherein said cell expresses said polypeptide, and b) determining whether or not said test molecule induces, in said cell, a mu3 opiate receptor-mediated response.
- 21. The method of claim 20, wherein said determining step comprises monitoring nitric oxide synthase activity in said cell.
- 22. The method of claim 21, wherein said monitoring nitric oxide synthase activity comprises detecting nitric oxide release from said cell.
- 23. The method of claim 22, wherein a nitric oxide-specific amperometric probe is used to detect said nitric oxide release.
- 24. The method of claim 20, wherein said determining step comprises monitoring intracellular calcium levels within said cell.
- 25. The method of claim 24, wherein a fluorescent ion indicator is used to monitor said intracellular calcium levels.
- 26. The method of claim 25, wherein said fluorescent ion indicator comprises Fura-2.
- 27. The method of claim 20, wherein said determining step comprises monitoring nitric oxide synthase activity and intracellular calcium levels in said cell.
- 28. A method for identifying a mu3 opiate receptor antagonist, said method comprising:
a) contacting a cell with a test molecule and a mu3 opiate receptor agonist, wherein said cell comprises an isolated nucleic acid molecule that encodes a polypeptide having mu3 opiate receptor activity, and wherein said cell expresses said polypeptide, and b) determining whether or not said test molecule reduces or prevents, in said cell, a mu3 opiate receptor-mediated response induced by said mu3 opiate receptor agonist.
- 29. The method of claim 28, wherein said mu3 opiate receptor agonist comprises morphine or dihydromorphine.
- 30. The method of claim 28, wherein said determining step comprises monitoring nitric oxide synthase activity in said cell.
- 31. The method of claim 28, wherein said determining step comprises monitoring intracellular calcium levels within said cell.
- 32. An isolated nucleic acid molecule comprising a nucleic acid sequence with a length and a percent identity to the sequence set forth in SEQ ID NO: 22 over said length, wherein the point defined by said length and said percent identity is within the area defined by points A, B, C, and D of FIG. 1, wherein point A has coordinates (225, 100), point B has coordinates (225, 65), point C has coordinates (15, 65), and point D has coordinates (15, 100).
CROSS-RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application Serial No. 60/270,479, filed Feb. 22, 2001 and U.S. Provisional Application Serial No. 60/336,677, filed Dec. 5, 2001.
STATEMENT AS TO FEDERALLY SPONSORED RESEARCH
[0002] Funding for the work described herein was provided by the federal government under NIDA number 5R24DA09010. Thus, the federal government may have certain rights in the invention.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60270479 |
Feb 2001 |
US |
|
60336677 |
Dec 2001 |
US |