Members of the capsaicin/vanilloid receptor family of proteins and uses thereof
The invention provides isolated nucleic acids molecules, designated hVR-1, hVR-2, and rVR-2 nucleic acid molecules, which encode novel members of the Capsaicin/Vanilloid receptor family. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing hVR-1, hVR-2, and rVR-2 nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which an hVR-1, hVR-2, and rVR-2 gene has been introduced or disrupted. The invention still further provides isolated hVR-1, hVR-2, and rVR-2 proteins, fusion proteins, antigenic peptides and anti-hVR-1, anti-hVR-2, and anti-rVR-2 antibodies. Diagnostic methods utilizing compositions of the invention are also provided.
1. A method for identifying a compound which binds to a polypeptide comprising the amino acid sequence of SEQ ID NO:2, the method comprising:
a) contacting a sample comprising the polypeptide with a test compound under conditions suitable for binding; and
b) determining whether the test compound binds to the polypeptide, thereby identifying a compound which binds to a polypeptide comprising the amino acid sequence of SEQ ID NO:2.
2. The method of claim 1 wherein the polypeptide consists of SEQ ID NO:2.
3. The method of claim 1 wherein the sample is a cell.
4. The method of claim 1 , wherein the polypeptide is isolated.
5. The method of claim 1 wherein the sample comprises a membrane-bound form of the polypeptide.
6. The method of claim 1 wherein binding of the test compound to the polypeptide is detected by the use of a direct binding assay.
7. The method of claim 1 wherein binding of the test compound to the polypeptide is detected by the use of a competition binding assay.
8. The method of claim 1 wherein binding of the test compound to the polypeptide is detected by the use of an assay for hVR-1 activity.
9. The method of claim 8 wherein the assay comprises:
(a) measuring in a first cell a first rapid inactivating inward current in the absence of a test compound;
(b) measuring in a second cell a second rapid inactivating inward current in the presence of the test compound, wherein the first and second rapid inactivating inward currents are stimulated by a temperature of 48-50° C. and wherein the first and second cells express the polypeptide comprising the amino acid sequence SEQ ID NO:2; and
(c) comparing the first and second rapid inactivating inward currents, wherein a difference between the first and second rapid inactivating inward currents identifies the test compound as a compound which modulates the activity of the polypeptide.
10. The method of claim 8 , wherein the hVR-1 activity is modulation of membrane depolarization.
11. The method of claim 8 , wherein the hVR-1 activity is modulation of intracellular calcium levels.
12. The method of claim 3 , wherein the cell is a neuronal cell.
13. The method of claim 9 wherein the first and second cells are neuronal cells.