Methods of assaying receptor activity and constructs useful in such methods
AssigneeDuke University
InventorsLawrence S. Barak, Marc G. Caron, Stephen S. Ferguson, Jie Zhang
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Abstract
Described are methods of detecting G-protein coupled receptor (GPCR) activity in vivo and in vitro; methods of assaying GPCR activity; and methods of screening for GPCR ligands, G Protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process. Constructs useful in such methods are described.
Claims26
A method of detecting G protein coupled receptor (GPCR) pathway activity in a cell expressing at least one GPCR and containing .beta.-arrestin protein conjugated to an optically detectable molecule, said method comprising detecting translocation of the detectable molecule from the cytosol of the cell to the membrane edge of the cell, wherein said translocation of the detectable molecule indicates activation of the GPCR pathway;
- and wherein said detecting step comprises (i) detecting an increase in said detectable molecule at said membrane edge;
- (ii) detecting a decrease in said detectable molecule in said cytosol;
- or (iii) detecting both an increase in said detectable molecule at said membrane edge and detecting a decrease in said detectable molecule in said cytosol.
Dependencies are inferred from the claim text.