Journal article
Identification of small-molecule inhibitors of RGS4 using a high-throughput flow cytometry protein interaction assay
Molecular pharmacology, Vol.71(1), pp.169-175
01/2007
DOI: 10.1124/mol.106.028670
PMID: 17012620
Abstract
Regulators of G-protein signaling (RGS) proteins are important components of signal transduction pathways initiated through G-protein-coupled receptors (GPCRs). RGS proteins accelerate the intrinsic GTPase activity of G-protein alpha-subunits (Galpha) and thus shorten the time course and reduce the magnitude of G-protein alpha- and betagamma-subunit signaling. Inhibiting RGS action has been proposed as a means to enhance the activity and specificity of GPCR agonist drugs, but pharmacological targeting of protein-protein interactions has typically been difficult. The aim of this project was to identify inhibitors of RGS4. Using a Luminex 96-well plate bead analyzer and a novel flow-cytometric protein interaction assay to assess Galpha-RGS interactions in a high-throughput screen, we identified the first small-molecule inhibitor of an RGS protein. Of 3028 compounds screened, 1, methyl N-[(4-chlorophenyl)sulfonyl]-4-nitrobenzenesulfinimidoate (CCG-4986), inhibited RGS4/Galpha(o) binding with 3 to 5 muM potency. It binds to RGS4, inhibits RGS4 stimulation of Galpha(o) GTPase activity in vitro, and prevents RGS4 regulation of mu-opioid-inhibited adenylyl cyclase activity in permeabilized cells. Furthermore, CCG-4986 is selective for RGS4 and does not inhibit RGS8. Thus, we demonstrate the feasibility of targeting RGS/Galpha protein-protein interactions with small molecules as a novel means to modulate GPCR-mediated signaling processes.
Details
- Title: Subtitle
- Identification of small-molecule inhibitors of RGS4 using a high-throughput flow cytometry protein interaction assay
- Creators
- David L Roman - Department of Pharmacology, University of Michigan Medical School, 1150 W. Medical Center Drive, 1303 MSRB III, Ann Arbor, MI 41809, USAJeffery N TalbotRebecca A RoofRoger K SunaharaJohn R TraynorRichard R Neubig
- Resource Type
- Journal article
- Publication Details
- Molecular pharmacology, Vol.71(1), pp.169-175
- Publisher
- United States
- DOI
- 10.1124/mol.106.028670
- PMID
- 17012620
- ISSN
- 0026-895X
- eISSN
- 1521-0111
- Grant note
- T32-DA007268 / NIDA NIH HHS DA004087 / NIDA NIH HHS F32-GM076821 / NIGMS NIH HHS GM039561 / NIGMS NIH HHS
- Language
- English
- Date published
- 01/2007
- Academic Unit
- Pharmacy; Iowa Neuroscience Institute; Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984065399402771
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