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US 7,741,064 B2Granted
Materials and methods relating to G-protein coupled receptor oligomers
AssigneeThe University Court of the University of Glasgow
InventorsGraeme Milligan, Dominic Behan
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Abstract
The invention provides materials and methods relating to G-protein coupled receptor (GPCR) oligomers. Complexes of two or more GPCRs associated with G-proteins are provided. Also provided are fusion proteins comprising a GPCR and a G-protein, nucleic acids, expression vectors and host cells. Methods of producing the complexes and fusion proteins of the invention are also provided.
Claims11
A GPCR oligomer comprising:
- (a) a first GPCR associated with a first G-protein wherein the first GPCR has a modified amino acid sequence compared to a wild-type GPCR sequence so as to render it non-functional with respect to the first G-protein;
- and;
- (b) a second GPCR associated with a second G-protein wherein the second G-protein is non-functional, wherein the wild-type form of the first GPCR is different to the second GPCR.
An isolated cell membrane comprising:
- (a) a first GPCR associated with a first G-protein wherein the first GPCR has a modified amino acid sequence compared to a wild-type GPCR sequence so as to render it non-functional with respect to the first G-protein;
- and,
- (b) a second GPCR associated with a second G-protein wherein the second G-protein is non-functional, wherein the wild-type form of the first GPCR is different to the second GPCR.
A eukaryotic cell comprising:
- a plasma membrane comprising:
- (a) a first GPCR associated with a first G-protein wherein the first GPCR has a modified amino acid sequence compared to a wild-type GPCR sequence so as to render it non-functional with respect to the first G-protein;
- and,
- (b) a second GPCR associated with a second G-protein wherein the second G-protein is non-functional, wherein the wild-type form of the first GPCR is different to the second GPCR.
1 more element
A method of making a cell comprising:
- (a) expressing a first nucleic acid construct in said cell, said nucleic acid construct encoding a first GPCR/G-protein fusion protein wherein the GPCR is mutated as compared to the wild-type GPCR thereby rendering it non-functional with respect to its G-protein;
- (b) expressing a second nucleic acid construct in said cell, said second nucleic acid construct encoding a second GPCR/G-protein fusion protein wherein the G-protein is mutated as compared to the wild-type G-protein thereby rendering it non-functional;
- and
- wherein the wild type form of the first GPCR is different to the second GPCR.
1 more element
A method of screening comprising:
- a) contacting a compound with a cell expressing a GPCR oligomer comprising (i) a first GPCR associated with a G-protein wherein the first GPCR has a modified amino acid sequence compared to the wild-type GPCR sequence so as to render it non-functional with respect to the first G-protein;
- and (ii) a second GPCR associated with a G-protein wherein the G-protein is modified so that it is non-functional, wherein the wild-type form of the first GPCR is different to the second GPCR;
- b) detecting the presence of a cellular signal resulting from contact between said compound and said GPCR oligomer;
- and
- c) determining an effect said compound has on said cellular signal.
2 more elements
Dependencies are inferred from the claim text.