G protein-coupled receptor internalization assays in the high-content screening format.

Abstract:

:High-content screening (HCS), a combination of fluorescence microscopic imaging and automated image analysis, has become a frequently applied tool to study test compound effects in cellular disease-modeling systems. This chapter describes the measurement of G protein-coupled receptor (GPCR) internalization in the HCS format using a high-throughput, confocal cellular imaging device. GPCRs are the most successful group of therapeutic targets on the pharmaceutical market. Accordingly, the search for compounds that interfere with GPCR function in a specific and selective way is a major focus of the pharmaceutical industry today. This chapter describes methods for the ligand-induced internalization of GPCRs labeled previously with either a fluorophore-conjugated ligand or an antibody directed against an N-terminal tag of the GPCR. Both labeling techniques produce robust assay formats. Complementary to other functional GPCR drug discovery assays, internalization assays enable a pharmacological analysis of test compounds. We conclude that GPCR internalization assays represent a valuable medium/high-throughput screening format to determine the cellular activity of GPCR ligands.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Haasen D,Schnapp A,Valler MJ,Heilker R

doi

10.1016/S0076-6879(06)14008-2

subject

Has Abstract

pub_date

2006-01-01 00:00:00

pages

121-39

eissn

0076-6879

issn

1557-7988

pii

S0076-6879(06)14008-2

journal_volume

414

pub_type

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