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Brenke, J.K. ; Salmina, E.S.* ; Ringelstetter, L. ; Dornauer, S. ; Kuzikov, M.* ; Rothenaigner, I. ; Schorpp, K.K. ; Giehler, F. ; Gopalakrishnan, J.* ; Kieser, A. ; Gul, S.* ; Tetko, I.V. ; Hadian, K.

Identification of small-molecule frequent hitters of glutathione S-transferase-glutathione interaction.

J. Biomol. Screen. 21, 596-607 (2016)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
In high-throughput screening (HTS) campaigns, the binding of glutathione S-transferase (GST) to glutathione (GSH) is used for detection of GST-tagged proteins in protein-protein interactions or enzyme assays. However, many false-positives, so-called frequent hitters (FH), arise that either prevent GST/GSH interaction or interfere with assay signal generation or detection. To identify GST-FH compounds, we analyzed the data of five independent AlphaScreen-based screening campaigns to classify compounds that inhibit the GST/GSH interaction. We identified 53 compounds affecting GST/GSH binding but not influencing His-tag/Ni(2+)-NTA interaction and general AlphaScreen signals. The structures of these 53 experimentally identified GST-FHs were analyzed in chemoinformatic studies to categorize substructural features that promote interference with GST/GSH binding. Here, we confirmed several existing chemoinformatic filters and more importantly extended them as well as added novel filters that specify compounds with anti-GST/GSH activity. Selected compounds were also tested using different antibody-based GST detection technologies and exhibited no interference clearly demonstrating specificity toward their GST/GSH interaction. Thus, these newly described GST-FH will further contribute to the identification of FH compounds containing promiscuous substructures. The developed filters were uploaded to the OCHEM website (http://ochem.eu) and are publicly accessible for analysis of future HTS results.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Alphascreen ; Gst ; Frequent Hitter ; Glutathione ; High-throughput Screening; Center-dot-s; Crystal-structures; Fusion Proteins; Drug Discovery; Purification; Inhibitors; Evolution; Chemicals; Database; Filters
ISSN (print) / ISBN 1087-0571
e-ISSN 1552-454X
Journal Journal of Biomolecular Screening
Quellenangaben Volume: 21, Issue: 6, Pages: 596-607 Article Number: , Supplement: ,
Publisher Sage
Publishing Place Thousand Oaks
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Molecular Toxicology and Pharmacology (TOXI)
Research Unit Gene Vector (AGV)
Institute of Structural Biology (STB)