PMID- 12769684
OWN - NLM
STAT- MEDLINE
DCOM- 20030805
LR  - 20190922
IS  - 1386-2073 (Print)
IS  - 1386-2073 (Linking)
VI  - 6
IP  - 4
DP  - 2003 Jun
TI  - Established and emerging fluorescence-based assays for G-protein function: 
      heterotrimeric G-protein alpha subunits and regulator of G-protein signaling 
      (RGS) proteins.
PG  - 399-407
AB  - 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 G-protein-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 G-protein alpha subunit regulation in vitro. Furthermore, we 
      consider the potential of such techniques in high-throughput screening and drug 
      discovery.
FAU - Kimple, Randall J
AU  - Kimple RJ
AD  - Department of Pharmacology, Lineberger Comprehensive Cancer Center, and UNC 
      Neuroscience Center, The University of North Carolina at Chapel Hill, Chapel 
      Hill, NC 27599 USA. dsiderov@med.unc.edu
FAU - Jones, Miller B
AU  - Jones MB
FAU - Shutes, Adam
AU  - Shutes A
FAU - Yerxa, Benjamin R
AU  - Yerxa BR
FAU - Siderovski, David P
AU  - Siderovski DP
FAU - Willard, Francis S
AU  - Willard FS
LA  - eng
GR  - F30 MH64319/MH/NIMH NIH HHS/United States
GR  - GM062338/GM/NIGMS NIH HHS/United States
GR  - GM065533/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United Arab Emirates
TA  - Comb Chem High Throughput Screen
JT  - Combinatorial chemistry & high throughput screening
JID - 9810948
RN  - 0 (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene)
RN  - 0 (Boron Compounds)
RN  - 0 (Protein Subunits)
RN  - 0 (RGS Proteins)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - EC 3.6.5.1 (Heterotrimeric GTP-Binding Proteins)
SB  - IM
MH  - Animals
MH  - Boron Compounds/chemistry
MH  - Fluorescence
MH  - Fluorescence Resonance Energy Transfer
MH  - Guanosine Triphosphate/metabolism
MH  - Heterotrimeric GTP-Binding Proteins/chemistry/*metabolism
MH  - Humans
MH  - Protein Subunits
MH  - RGS Proteins/chemistry/*metabolism
MH  - Signal Transduction
MH  - Spectrometry, Fluorescence/methods
EDAT- 2003/05/29 05:00
MHDA- 2003/08/06 05:00
CRDT- 2003/05/29 05:00
PHST- 2003/05/29 05:00 [pubmed]
PHST- 2003/08/06 05:00 [medline]
PHST- 2003/05/29 05:00 [entrez]
AID - 10.2174/138620703106298491 [doi]
PST - ppublish
SO  - Comb Chem High Throughput Screen. 2003 Jun;6(4):399-407. doi: 
      10.2174/138620703106298491.