G protein-coupled receptor fusion proteins in drug discovery.

Abstract:

:A wide range of peptides and polypeptides can be appended to either the N- or C-terminus of G protein-coupled receptors without disrupting substantially ligand binding and signal transduction. Following fusion of fluorescent proteins, reporter gene constructs or G protein alpha subunits to the C-terminal tail of a receptor high content and G protein activation assays can be employed to identify agonist ligands. Further modification of the receptor fusions to introduce enhanced levels of constitutive activity and to physically destabilise the protein allows antagonist/inverse agonists screens to be developed in parallel. Equivalent C-terminal addition of pairs of complementary, non-functional, polypeptide fragments allows the application of enzyme complementation techniques. Introduction of N-terminal tags to receptors has also allowed the introduction of novel assay techniques based on a pH-sensitive cyanine dye. These have the capacity to overcome certain limitations of GPCR-fluorescent protein fusions.

journal_name

Curr Pharm Des

authors

Milligan G,Feng GJ,Ward RJ,Sartania N,Ramsay D,McLean AJ,Carrillo JJ

doi

10.2174/1381612043384295

subject

Has Abstract

pub_date

2004-01-01 00:00:00

pages

1989-2001

issue

17

eissn

1381-6128

issn

1873-4286

journal_volume

10

pub_type

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