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. An isolated feline TAS2R (fTAS2R) receptor polypeptide comprising the sequence SEQ ID NO:14, wherein the polypeptide is covalently bound to a label, a solid support, a lipid monolayer, or a heterologous polypeptide.
2. The polypeptide of claim 1 , wherein the polypeptide is covalently bound to a heterologous polypeptide.
3. The polypeptide of claim 2 , wherein the heterologous polypeptide is covalently bound to the amino terminus or the carboxy terminus of the feline TAS2R receptor polypeptide.
4. A method for identifying a compound that interacts with a feline TAS2R receptor polypeptide comprising:
contacting the polypeptide of claim 1 with a test compound, and
detecting interaction between the polypeptide and the test compound.
5. A method for identifying a compound which modulates a feline TAS2R receptor polypeptide which comprises:
contacting the polypeptide of claim 1 with a TAS2R receptor ligand in both the presence and absence of a test compound in separate assays, and
determining whether the test compound modulates binding of the ligand to the receptor polypeptide or activation of the receptor polypeptide by the ligand.
6. The method of claim 5 , wherein the polypeptide is bound to a solid support, expressed in a host cell, in a bilayer membrane, in a lipid monolayer, or in a vesicle.
7. A method of preparing an edible composition comprising
contacting an edible composition or a component thereof with the polypeptide of claim 1 for a time sufficient to reduce the amount of a bitter compound in the edible composition or component thereof.
8. The method of claim 7 wherein the polypeptide is bound to a solid support that can be separated from the edible composition.
9. The method of claim 7 wherein the edible composition is a feline food composition.
10. The method of claim 4 , wherein the polypeptide is bound to a solid support, expressed in a host cell, in a bilayer membrane, in a lipid monolayer, or in a vesicle.
11. The method of claim 4 , wherein detecting interaction between the polypeptide and the test compound comprises
measuring an electrical property, measuring a change in an ion concentration, measuring a change in protein conformation, measuring binding of the test compound to the polypeptide, measuring a change in phosphorylation level, measuring a change in transcription level, measuring a change in second messenger level, measuring a change in neurotransmitter level, measuring a change in a spectroscopic characteristic, measuring a change in a hydrodynamic property, measuring a change in a chromatographic property, or measuring a change in solubility.
12. The method of claim 5 , wherein determining whether the test compound modulates binding of the ligand to the receptor or activation of the receptor by the ligand comprises
measuring an electrical property, measuring an ion concentration, measuring a change in protein conformation, measuring a binding of the test compound to the polypeptide, measuring a change in phosphorylation level, measuring a change in transcription level, measuring a change in second messenger level, or measuring a change in neurotransmitter level.
13. The method of claim 7 , further comprising
contacting the edible composition or the component thereof with a peptide comprising a sequence selected from SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26.
14. The polypeptide of claim 1 , wherein the polypeptide is covalently bound to a label.
15. The polypeptide of claim 1 , wherein the polypeptide is covalently bound to a solid support.
16. The polypeptide of claim 1 , wherein the polypeptide is covalently bound to a lipid monolayer.