Feline bitter taste receptors and methods
View Patent ↗A family of novel feline bitter taste receptors, referred to as feline TAS2R (fTAS2R), are disclosed herein. Isolated polynucleotides encoding the novel feline bitter taste receptors and chimeric polypeptides are also disclosed, as are expression vectors and host cells for expression of the novel feline bitter taste receptors. Methods of identifying compounds that bind to the novel feline bitter taste receptors and modulate their activity are disclosed.
1. A polypeptide comprising
the amino acid sequence of SEQ ID NO:16, or
an amino acid sequence having at least 95% homology to the sequence of SEQ ID NO:16;
wherein the polypeptide is covalently bound to a label, a solid support, or a lipid monolayer.
2. The polypeptide of claim 1 comprising the amino acid sequence of SEQ ID NO:16.
3. The polypeptide of claim 1 comprising an amino acid sequence having at least 95% homology to SEQ ID NO:16.
4. The polypeptide of claim 1 , wherein the polypeptide is covalently bound to a lipid monolayer.
5. The polypeptide of claim 1 , wherein the polypeptide is covalently bound to a label.
6. The polypeptide of claim 5 , wherein the label is a heterologous polypeptide sequence.
7. The polypeptide of claim 6 , wherein the heterologous polypeptide sequence is a membrane targeting sequence or a peptide tag sequence.
8. The polypeptide of claim 5 , wherein the label is a radiolabel, a fluorescence label, a chemiluminescent label, an enzymic label, or an immunogenic label.
9. The polypeptide of claim 1 , wherein the polypeptide is covalently bound to a solid support.
10. A chimeric polypeptide comprising at least one extracellular domain of SEQ ID NO:16 covalently linked to a heterologous polypeptide sequence, wherein the extracellular domain comprises amino acids 1-8, 69-88, 150-185, or 252-261 of SEQ ID NO:16.
11. A chimeric polypeptide comprising at least one intracellular domain of SEQ ID NO:16 covalently linked to a heterologous polypeptide sequence, wherein the intracellular domain comprises amino acids 30-47, 110-128, 207-230, or 283-309 of SEQ ID NO:16.
12. A chimeric polypeptide comprising at least one transmembrane domain of SEQ ID NO:16 covalently linked to a heterologous polypeptide sequence, wherein the transmembrane domain comprises amino acids 9-29, 48-68, 89-109, 129-149, 186-206, 231-251, or 262-282 of SEQ ID NO:16.
13. The chimeric polypeptide of claim 12 comprising the transmembrane domains comprising amino acids 89-109, 231-251, and 262-282 of SEQ ID NO:16.
14. The chimeric polypeptide of claim 12 comprising the transmembrane domains comprising amino acids 89-109, 186-206, 231-251, and 262-282 of SEQ ID NO:16.
15. The chimeric polypeptide of claim 12 comprising the transmembrane domains comprising amino acids 89-109, 129-149, 186-206, 231-251, and 262-282 of SEQ ID NO:16.
16. The chimeric polypeptide of claim 12 comprising the transmembrane domains comprising amino acids 9-29, 48-68, 89-109, 129-149, 186-206, 231-251, and 262-282 of SEQ ID NO:16.
17. The chimeric polypeptide of claim 16 further comprising amino acids 69-88 of SEQ ID NO:16.
18. A method for identifying a compound that interacts with a feline TAS2R receptor comprising:
contacting the polypeptide of claim 1 with a test compound, and
detecting interaction between the polypeptide and the test compound.
19. The method of claim 18 , wherein the polypeptide is bound to a solid support or expressed in a host cell.
20. A method for identifying a compound which modulates a feline TAS2R receptor which comprises:
contacting a polypeptide of claim 1 with a receptor ligand in the presence or absence of a test compound, and
determining whether the test compound modulates binding of the ligand to the polypeptide or activation of the polypeptide by the ligand.