A covalent peptide inhibitor of RGS4 identified in a focused one-bead, one compound library screen

Rebecca A Roof; David L Roman; Samuel T Clements; Katarzyna Sobczyk-Kojiro; Levi L Blazer; Shodai Ota; Henry I Mosberg; Richard R Neubig

doi:10.1186/1471-2210-9-9

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A covalent peptide inhibitor of RGS4 identified in a focused one-bead, one compound library screen

Journal article

Open access

Peer reviewed

A covalent peptide inhibitor of RGS4 identified in a focused one-bead, one compound library screen

Rebecca A Roof, David L Roman, Samuel T Clements, Katarzyna Sobczyk-Kojiro, Levi L Blazer, Shodai Ota, Henry I Mosberg and Richard R Neubig

BMC pharmacology, Vol.9(1), pp.9-9

05/22/2009

DOI: 10.1186/1471-2210-9-9

PMCID: PMC2700083

PMID: 19463173

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Abstract

Regulators of G protein signaling (RGSs) accelerate GTP hydrolysis by Galpha subunits and profoundly inhibit signaling by G protein-coupled receptors (GPCRs). The distinct expression patterns and pathophysiologic regulation of RGS proteins suggest that inhibitors may have therapeutic potential. We recently described a focused one-bead, one-compound (OBOC) library screen to identify peptide inhibitors of RGS4. Here we extend our observations to include another peptide with a different mechanism of action. Peptide 5nd (Tyr-Trp-c [Cys-Lys-Gly-Leu-Cys]-Lys-NH2, S-S) blocks the RGS4-Galphao interaction with an IC50 of 28 microM. It forms a covalent, dithiothreitol (DTT) sensitive adduct with a mass consistent with the incorporation of one peptide per RGS. Peptide 5nd activity is abolished by either changing its disulfide bridge to a methylene dithioether bridge, which cannot form disulfide bridges to the RGS, or by removing all cysteines from the RGS protein. However, no single cysteine in RGS4 is completely necessary or sufficient for 5nd activity. Though it has some RGS selectivity, 5nd appears to be a partially random cysteine modifier. These data suggest that it inhibits RGS4 by forming disulfide bridges with the protein.

Amino Acid Sequence

Peptides, Cyclic - chemical synthesis

RGS Proteins - chemistry

Molecular Sequence Data

Peptides, Cyclic - pharmacology

Protein Conformation

RGS Proteins - antagonists & inhibitors

Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Cysteine - chemistry

Peptide Library

Peptides, Cyclic - chemistry

Details

Title: Subtitle: A covalent peptide inhibitor of RGS4 identified in a focused one-bead, one compound library screen
Creators: Rebecca A Roof - Department of Pharmacology, University of Michigan,1301 MSRB III SPC 5632, 1150 W, Medical Center Dr, Ann Arbor, MI 48109, USA. roofra@ninds.nih.gov
David L Roman
Samuel T Clements
Katarzyna Sobczyk-Kojiro
Levi L Blazer
Shodai Ota
Henry I Mosberg
Richard R Neubig
Resource Type: Journal article
Publication Details: BMC pharmacology, Vol.9(1), pp.9-9
DOI: 10.1186/1471-2210-9-9
PMID: 19463173
PMCID: PMC2700083
NLM abbreviation: BMC Pharmacol
ISSN: 1471-2210
eISSN: 1471-2210
Publisher: England
Grant note: T32 GM00008597 / NIGMS NIH HHS P30 CA046592 / NCI NIH HHS GM39561 / NIGMS NIH HHS R01 DA003910 / NIDA NIH HHS P30 DK020572 / NIDDK NIH HHS P60 DK020572 / NIDDK NIH HHS P30CA046592 / NCI NIH HHS DA003910 / NIDA NIH HHS T32 GM008597 / NIGMS NIH HHS DA23252 / NIDA NIH HHS DK20572 / NIDDK NIH HHS R01 GM039561 / NIGMS NIH HHS R01 DA023252 / NIDA NIH HHS
Language: English
Date published: 05/22/2009
Academic Unit: Pharmacy; Iowa Neuroscience Institute; Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
Record Identifier: 9984065489002771

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