Development of a novel high-throughput screen and identification of small-molecule inhibitors of the Gα-RGS17 protein-protein interaction using AlphaScreen

Duncan I Mackie; David L Roman

doi:10.1177/1087057111410427

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Development of a novel high-throughput screen and identification of small-molecule inhibitors of the Gα-RGS17 protein-protein interaction using AlphaScreen

Journal article

Open access

Development of a novel high-throughput screen and identification of small-molecule inhibitors of the Gα-RGS17 protein-protein interaction using AlphaScreen

Duncan I Mackie and David L Roman

Journal of biomolecular screening, Vol.16(8), pp.869-877

09/2011

DOI: 10.1177/1087057111410427

PMCID: PMC5525514

PMID: 21680864

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Abstract

In this study, the authors used AlphaScreen technology to develop a high-throughput screening method for interrogating small-molecule libraries for inhibitors of the Gα(o)-RGS17 interaction. RGS17 is implicated in the growth, proliferation, metastasis, and the migration of prostate and lung cancers. RGS17 is upregulated in lung and prostate tumors up to a 13-fold increase over patient-matched normal tissues. Studies show RGS17 knockdown inhibits colony formation and decreases tumorigenesis in nude mice. The screen in this study uses a measurement of the Gα(o)-RGS17 protein-protein interaction, with an excellent Z score exceeding 0.73, a signal-to-noise ratio >70, and a screening time of 1100 compounds per hour. The authors screened the NCI Diversity Set II and determined 35 initial hits, of which 16 were confirmed after screening against controls. The 16 compounds exhibited IC(50) <10 µM in dose-response experiments. Four exhibited IC(50) values <6 µM while inhibiting the Gα(o)-RGS17 interaction >50% when compared to a biotinylated glutathione-S-transferase control. This report describes the first high-throughput screen for RGS17 inhibitors, as well as a novel paradigm adaptable to many other RGS proteins, which are emerging as attractive drug targets for modulating G-protein-coupled receptor signaling.

Binding, Competitive

Recombinant Proteins - metabolism

Lung Neoplasms - drug therapy

Small Molecule Libraries - analysis

Small Molecule Libraries - pharmacology

Recombinant Proteins - antagonists & inhibitors

Escherichia coli

GTP-Binding Proteins - antagonists & inhibitors

Humans

Signal-To-Noise Ratio

Male

Dose-Response Relationship, Drug

Small Molecule Libraries - chemistry

High-Throughput Screening Assays

Transformation, Bacterial

Plasmids

Protein Binding - drug effects

RGS Proteins - metabolism

RGS Proteins - antagonists & inhibitors

Prostatic Neoplasms - drug therapy

GTP-Binding Proteins - metabolism

Details

Title: Subtitle: Development of a novel high-throughput screen and identification of small-molecule inhibitors of the Gα-RGS17 protein-protein interaction using AlphaScreen
Creators: Duncan I Mackie - Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, 115 South Grand Avenue, Iowa City, IA 52242, USA
David L Roman
Resource Type: Journal article
Publication Details: Journal of biomolecular screening, Vol.16(8), pp.869-877
Publisher: United States
DOI: 10.1177/1087057111410427
PMID: 21680864
PMCID: PMC5525514
ISSN: 1087-0571
eISSN: 1552-454X
Grant note: T32 GM008365 / NIGMS NIH HHS
Language: English
Date published: 09/2011
Academic Unit: Pharmacy; Iowa Neuroscience Institute; Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
Record Identifier: 9984065382102771

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