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Identification of novel inhibitors of RGS6 through an alpha screen highthroughput campaign (843.14)
Author(s) -
Bodle Christopher,
Fisher Rory,
Roman David
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.843.14
Subject(s) - heterotrimeric g protein , regulator of g protein signaling , g protein coupled receptor , gtpase activating protein , g protein , chemistry , gene knockdown , biology , microbiology and biotechnology , gtpase , signal transduction , biochemistry , gene
G Protein Coupled Receptors (GPCRs) are highly targeted proteins for drug discovery, with over 50% of all drugs on the market targeting these receptors. Regulator of G Protein Signaling (RGS) Proteins regulate the signaling cascade of GPCRs by accelerating the intrinsic GTPase activity of G protein alpha subunits and returning the heterotrimeric G protein to its inactive state. Larger RGS proteins can regulate a number of ancillary signaling cascades as well through additional biding motifs. RGS proteins have emerged as attractive targets for drug design, both by targeting their GTPase dependent and independent activity. RGS6 is one such RGS protein. RGS6 has been shown to be important in several disease states. Our lab employed alpha screen technology to identify novel inhibitors of RGS6 by screening for compounds which disrupt the protein:protein interaction between Gα o and the RH domain of RGS6. The RH domain of RGS6 was purified largely as described previously, and its activity was determined via a Malachite Green free phosphate detection assay. Assay robustness was determined resulting in a z‐score of 0.60. The screen yielded 5 compounds which inhibited the RGS6 RH : Gα o interaction at greater than 40%. Dose response curves were determined which resulted in three compounds with double‐digit micromolar IC 50 ’s.