Gustducin gamma subunit materials and methods
AssigneeMount Sinai School of Medicine
InventorsLiquan Huang, Robert F. Margolskee, Marianna Max, Y. Gopi Shanker
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Abstract
Gustducin is a taste receptor cell heterotrimeric guanine nucleotide binding protein. Disclosed herein is the amino acid sequence of the gamma (γ) subunit of gustducin, as well as polynucleotide sequences encoding the γ subunit. Also disclosed are methods of modifying taste involving agents that inhibit or activate the gustducin γ subunit, methods for identifying such taste modifying agents and various taste modifying agents.
Claims14
A purified and isolated polynucleotide sequence encoding gustducin gamma (γ) subunit polypeptide or a fragment or a variant thereof selected from the group consisting of:
- a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:
- 6;
- b) a polynucleotide encoding the amino acid sequence of SEQ ID NO:
- 7;
- c) a polynucleotide that hybridizes under moderately stringent conditions to the complement of the polynucleotide of a).
Purified and isolated gustducin γ subunit polypeptide, fragment or variant possessing at least one binding activity or immunological property specific to the gustducin γ subunit.
An antibody substance specific for the gustducin γ subunit.
A method for identifying a taste modifying agent comprising the steps of a) incubating phospholipid vesicles having gustducin γ subunit associated in biologically active form with a G protein β subunit and gustducin β subunit or transducin β subunit, b) adding radioactively labeled GTPγS in the presence and absence of an agent, and c) measuring the rate of GTPγS binding by the α subunit in the presence of the agent compared to the rate in the absence of the agent, wherein an increase in the rate of binding indicates that the agent is a taste stimulator and a decrease in the rate of binding indicates that the agent is a taste inhibitor.
A method for identifying a taste modifying agent comprising the steps of a) incubating phospholipid vesicles having gustducin γ subunit associated in biologically active form with a G protein β subunit and gustducin β subunit or transducin α subunit, b) adding GTP in the presence and absence of an agent, and c) measuring the rate of conversion of GTP to GDP by the α subunit in the presence of the agent compared to the rate in the absence of the agent, wherein an increase in the rate of conversion indicates that the agent is a taste stimulator and a decrease in the rate of conversion indicates that the agent is a taste inhibitor.
A method for identifying a taste modifying agent comprising the steps of a) incubating activated gustducin γ associated in biologically active form with a G protein β subunit and phospholipase C, b) adding 3H-myoinositol in the presence and absence of an agent, and c) measuring inositol trisphosphate generation in the presence of the agent in comparison to in the absence of the agent, wherein an increase in inositol trisphosphate generation indicates the agent is a taste stimulator and a decrease in inositol trisphosphate generation indicates that the agent is a taste inhibitor.
A method for modifying taste comprising the step of delivering to taste receptor cells a taste modifying agent selected from the group consisting of:
- a) a peptide comprising about amino acid 1 through about amino acid 14 of SEQ ID NO:
- 7;
- b) a peptide comprising about amino acid 18 through about amino acid 32 of SEQ ID NO:
- 7;
- c) a peptide comprising about amino acid 31 through about amino acid 40 of SEQ ID NO:
- 7;
- d) a peptide comprising about amino acid 47 through about amino acid 59 of SEQ ID NO:
- 7;
- and
- e) peptides comprising about amino acid 56 through about amino acid 67 of SEQ ID NO:
- 7.
Dependencies are inferred from the claim text.