Identification of Small-Molecule Frequent Hitters of Glutathione S-Transferase-Glutathione Interaction.

Autor:	Brenke JK; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Neuherberg, Germany., Salmina ES; Institute for Organic Chemistry, Technical University Bergakademie Freiberg, Germany., Ringelstetter L; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Neuherberg, Germany., Dornauer S; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Neuherberg, Germany., Kuzikov M; Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Hamburg, Germany., Rothenaigner I; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Neuherberg, Germany., Schorpp K; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Neuherberg, Germany., Giehler F; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Research Unit Gene Vectors, Munich, Germany German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany., Gopalakrishnan J; Laboratory for Centrosome and Cytoskeleton Biology, CMMC, Cologne, Germany., Kieser A; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Research Unit Gene Vectors, Munich, Germany German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany., Gul S; Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Hamburg, Germany., Tetko IV; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Institute of Structural Biology, Neuherberg, Germany BigChem GmbH, Ingolstädter Landstrasse 1, Neuherberg, Germany., Hadian K; Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Neuherberg, Germany kamyar.hadian@helmholtz-muenchen.de.
Jazyk:	angličtina
Zdroj:	Journal of biomolecular screening [J Biomol Screen] 2016 Jul; Vol. 21 (6), pp. 596-607. Date of Electronic Publication: 2016 Apr 04.
DOI:	10.1177/1087057116639992
Abstrakt:	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. (© 2016 Society for Laboratory Automation and Screening.)
Databáze:	MEDLINE
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