Mammalian sweet and amino acid heterodimeric taste receptors comprising T1R3 and T1R1
View Patent ↗The present invention provides isolated nucleic acid and amino acid sequences of sweet or amino acid taste receptors comprising T1R3 and T1R1, two heterologous G-protein coupled receptor polypeptides from the T1R family of sensory G-protein coupled receptors, antibodies to such receptors, methods of detecting such nucleic acids and receptors, and methods of screening for modulators of sweet and amino acid taste receptors.
1. An isolated receptor, wherein the receptor comprises a T1R3 polypeptide and a heterologous polypeptide, wherein the T1R3 polypeptide has at least 94% amino acid sequence identity to SEQ ID NO:25 and the heterologous polypeptide is a T1R1 polypeptide that has at least 90% amino acid sequence identity to SEQ ID NO:1.
2. The isolated receptor of claim 1 , wherein the receptor is a taste receptor.
3. The isolated receptor of claim 1 , wherein the T1R3 polypeptide has the amino acid sequence of SEQ ID NO:25.
4. The isolated receptor of claim 1 , wherein the T1R1 polypeptide has the amino acid sequence of SEQ ID NO:1.
5. The isolated receptor of claim 1 , wherein the T1R3 polypeptide is recombinant.
6. A host cell comprising the isolated receptor of claim 5 .
7. The isolated receptor of claim 1 , wherein the T1R3 polypeptide and the heterologous polypeptide are recombinant.
8. A host cell expressing the recombinant receptor of claim 7 .
9. The isolated receptor of claim 1 , wherein the T1R1 polypeptide is recombinant.
10. A host cell expressing the recombinant receptor of claim 9 .
11. The isolated receptor of claim 1 , wherein the T1R3 polypeptide and the heterologous polypeptide are non-covalently linked.
12. The isolated receptor of claim 1 , wherein the T1R3 polypeptide and the heterologous polypeptide are covalently linked.
13. The isolated receptor of claim 1 , wherein the receptor binds to at least one amino acid taste ligand.
14. The isolated receptor of claim 13 , wherein the taste ligand is selected from the group consisting of cysteine, methionine, arginine, valine, aspartic acid, glutamic acid, lysine, proline, leucine, isoleucine, alanine, asparagine, histidine, phenylalanine, tryptophan, glutamine, serine, threonine, glycine, glutamate, mono-sodium glutamate, and L-AP4.
15. The isolated receptor of claim 1 , wherein the receptor specifically binds to an antibody that specifically binds to a polypeptide comprising SEQ ID NO:25 or SEQ ID NO:1.
16. A method of identifying a compound that activates or inhibits taste signal transduction in taste cells, the method comprising the steps of:
(i) contacting the compound with a cell expressing the taste receptor of claim 1 in the plasma membrane of the cell; and
(ii) determining a functional effect of the compound upon the receptor, wherein said functional effect is binding to the receptor or having an effect on the activity of the receptor, thereby identifying a compound that activates or inhibits taste signal transduction.
17. The method of claim 16 , wherein the T1R3 polypeptide has the amino acid sequence of SEQ ID NO:25.
18. The method of claim 16 , wherein the T1R1 polypeptide has the amino acid sequence of SEQ ID NO:1.
19. The method of claim 16 , wherein the functional effect is determined by measuring binding of the compound to the receptor.
20. The method of claim 16 , wherein the functional effect is determined by measuring ligand binding to the receptor.
21. The method of claim 16 , wherein the functional effect is a chemical or phenotypic effect.
22. The method of claim 16 , wherein the functional effect is determined by measuring changes in intracellular cAMP, IP3, or Ca2+.
23. The method of claim 16 , wherein the cell is a mammalian cell.
24. The method of claim 23 , wherein the cell is a human cell.
25. The method of claim 16 , wherein the T1 R3 polypeptide and the T1 R1 polypeptide are non-covalently linked.
26. The method of claim 16 , wherein the T1 R3 polypeptide and the T1 R1 polypeptide are covalently linked.
27. The method of claim 16 , wherein the T1 R3 and T1 R1 polypeptides are both recombinant.
28. A method of identifying a compound that activates or inhibits taste signal transduction in taste cells, the method comprising the steps of: (i) contacting the compound with a cell expressing the taste receptor of claim 1 in the plasma membrane of the cell, wherein the T1 R3 polypeptide has the amino acid sequence of SEQ ID NO:25 and the T1 R1 polypeptide has the amino acid sequence of SEQ ID NO:1 ; and (ii) determining a functional effect of the compound upon the receptor, wherein said functional effect is binding to the receptor or having an effect on the activity of the receptor, thereby identifying a compound that activates or inhibits taste signal transduction, wherein the receptor specifically binds an amino acid, and at least one of the T1 R3 and T1 R1 polypeptides is recombinant.