Methods for identifying compounds which modulate T2R bitter taste receptors
The invention provides isolated nucleic acid and amino acid sequences of taste cell specific G-protein coupled receptors, antibodies to such receptors, methods of detecting such nucleic acids and receptors, and methods of screening for modulators of taste cell specific G-protein coupled receptors.
1. A method for identifying a compound that modulates taste signaling in taste cells, the method comprising the steps of:
(i) contacting the compound with a taste transduction G-protein coupled receptor polypeptide, wherein the polypeptide is expressed in a taste cell, the polypeptide comprising greater than 90% amino acid sequence identity to SEQ ID NO:60, wherein the receptor polypeptide binds bitter tastants; and
(ii) determining the functional effect of the compound upon the polypeptide, wherein said effect identifies a compound which modulates taste signaling in taste cells.
2. The method of claim 1 , wherein the polypeptide has G-protein coupled receptor activity.
3. The method of claim 1 , wherein the functional effect is determined by measuring changes in intracellular cAMP, cGMP, IP3, or Ca2+.
4. The method of claim 1 , wherein he functional effect is determined by measuring binding of the compound to an extracellular domain of the polypeptide.
5. The method of claim 1 , wherein the functional effect is determined by measuring binding of radiolabeled GTP to the polypeptide.
6. The method of claim 1 , wherein the polypeptide is recombinant.
7. The method of claim 1 , wherein the functional effect is measured by determining changes in the electrical activity of a cell expressing the polypeptide.
8. The method of claim 1 , wherein the cell is a eukaryotic cell.
9. The method of claim 1 , wherein the polypeptide comprises greater than 95% amino acid sequence identity to SEQ ID NO:60.
10. The method of claim 9 , wherein the polypeptide has G-protein coupled receptor activity.
11. The method of claim 9 , wherein the functional effect is determined by measuring changes in intracellular cAMP, cGMP, IP3, or Ca2+.
12. The method of claim 9 , wherein the functional effect is determined by measuring binding of the compound to an extracellular domain of the polypeptide.
13. The method of claim 9 , wherein the functional effect is determined by measuring binding of radiolabeled GTP to the polypeptide.
14. The method of claim 9 , wherein the polypeptide is recombinant.
15. The method of claim 9 , wherein the functional effect is measured by determining changes in the electrical activity of a cell expressing the polypeptide.
16. The method of claim 9 , wherein the cell is a eukaryotic cell.
17. The method of claim 1 , wherein the polypeptide comprises 100% amino acid identity to SEQ ID NO:60.