Mammalian sweet and amino acid heterodimeric taste receptors comprising T1R3 and T1R1

Patent metadata

Field	Value
Publication number	US-9347935-B2
Application number	US-201313903820-A
Country	US
Kind code	B2
Filing date	May 28, 2013
Priority date	Jul 28, 1998
Publication date	May 24, 2016
Grant date	May 24, 2016

Abstract

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.

First claim

What is claimed is:

1. A method for determining a functional effect of a compound upon a cell expressing a first recombinant polypeptide and a second recombinant polypeptide, the method comprising the steps of:
(a) expressing the first recombinant polypeptide and the second recombinant polypeptide in a plasma membrane of the cell,
wherein the first recombinant polypeptide comprises a first extracellular region and a first transmembrane region, wherein:
(i) the first extracellular region amino acid sequence is selected from the group consisting of: an amino acid sequence that has at least 93% amino acid sequence identity to the extracellular region of SEQ ID NO:20 or an amino acid sequence that has at least 94% amino acid sequence identity to the extracellular region of SEQ ID NO:23,
(ii) the first transmembrane region amino acid sequence is selected from the group consisting of: an amino acid sequence that has at least 93% amino acid sequence identity to the transmembrane region of SEQ ID NO:20 or has at least 94% amino acid sequence identity to the transmembrane region of SEQ ID NO:23, and
the second recombinant polypeptide comprises a second extracellular region and a second transmembrane region, and wherein the second extracellular region has an amino acid sequence having at least 90% amino acid sequence identity to the extracellular region of SEQ ID NO:2 and the second transmembrane region has an amino acid sequence having at least 90% amino acid sequence identity to the transmembrane region of SEQ ID NO:2,

(b) assaying a parameter in the cell, the parameter indirectly or directly under the influence of a T1R GPCR protein or a sweet and/or an amino acid taste receptor comprising one or more T1R GPCR proteins wherein the parameter is ligand binding or transduction of a cellular signal in response to ligand binding;
(c) contacting the cell with the compound;
(d) assaying the cell for an increase or decrease in the parameter, thereby determining the functional effect, if any, of the compound.

2. The method of claim 1 , wherein the extracellular region of the first polypeptide has at least 93% amino acid sequence identity to the extracellular region of SEQ ID NO:20 and the transmembrane region of the first polypeptide has at least 94% amino acid sequence identity to the transmembrane region of SEQ ID NO:23.

3. The method of claim 1 , wherein the transmembrane region of the first polypeptide has at least 93% amino acid sequence identity to the transmembrane region of SEQ ID NO:20 and the extracellular region of the first polypeptide has at least 94% amino acid sequence identity to the extracellular region of SEQ ID NO:23.

4. The method of claim 1 , wherein the first polypeptide comprises the extracellular region of SEQ ID NO:23 and the transmembrane region of SEQ ID NO:20.

5. The method of claim 1 , wherein the first polypeptide comprises the extracellular region of SEQ ID NO:20 and the transmembrane region of SEQ ID NO:23.

6. The method of claim 1 , wherein the extracellular region of the second polypeptide has the amino acid sequence of the extracellular region of SEQ ID NO:2.

7. The method of claim 1 , wherein the transmembrane region of the second polypeptide has the amino acid sequence of the transmembrane region of SEQ ID NO:2.

8. The method of claim 1 , wherein the first polypeptide and the second polypeptide are covalently linked.

9. The method of claim 1 , wherein the functional effect is determined by measuring ligand binding to cell expressing the first polypeptide and the second polypeptide.

10. The method of claim 1 , wherein the functional effect is a chemical or phenotypic effect.

11. The method of claim 1 , wherein the functional effect is determined by measuring changes in intracellular cAMP, IP3 or Ca 2+ .

12. The method of claim 1 , wherein the cell is a mammalian cell.

13. The method of claim 1 , wherein the cell is a human cell.

Assignees

• Univ California
• Us Health

Inventors

• Zuker Charles S
• Chandrashekar Jayaram
• Nelson Greg
• Zhang Yifeng
• Ryba Nicholas J P
• Hoon Mark A

Classifications

• C07K14/705 Primary

  Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} · CPC title

• G01N33/5076 Primary

  involving cell organelles, e.g. Golgi complex, endoplasmic reticulum · CPC title