Fluorescence-Based Assays for G-Protein (archive)
Established and Emerging Fluorescence-Based Assays for G-Protein
Function: Heterotrimeric G-Protein Alpha Subunits and Regulator of GProtein
Signaling (RGS) Proteins. (pdf file)
Randall J., et al. Combinatorial Chemistry & High Throughput Screening, 2003, 6.
Abstract: Heterotrimeric G-proteins are molecular switches that couple serpentine receptors to intracellular
effector pathways and the regulation of cell physiology. Ligand-bound receptors cause G-protein alpha
subunits to bind guanosine 5'-triphosphate (GTP) and activate effector pathways. Signal termination is
facilitated by the intrinsic GTPase activity of G-protein alpha subunits. Regulators of G-protein signaling
(RGS) proteins accelerate the GTPase activity of the G-protein alpha subunit, and thus negatively regulate Gprotein-
mediated signal transduction. In vitro biochemical assays of heterotrimeric G-proteins commonly
include measurements of nucleotide binding, GTPase activity, and interaction with RGS proteins. However, the
conventional assays for most of these processes involve radiolabeled guanine nucleotide analogues and scintillation counting. In this article, we focus on fluorescence-based methodologies to study heterotrimeric Gprotein alpha subunit regulation in vitro. Furthermore, we consider the potential of such techniques in highthroughput
screening and drug discovery.
Last update 21-Jan-2005, Rating n/a of 0 votes.
