Universität zu Köln

Brenke, Jara K., Salmina, Elena S., Ringelstetter, Larissa, Dornauer, Scarlett, Kuzikov, Maria, Rothenaigner, Ina, Schorpp, Kenji, Giehler, Fabian, Gopalakrishnan, Jay, Kieser, Arnd ORCID: 0000-0003-0783-1950, Gul, Sheraz, Tetko, Igor V. ORCID: 0000-0002-6855-0012 and Hadian, Kamyar ORCID: 0000-0001-8727-2575 (2016). Identification of Small-Molecule Frequent Hitters of Glutathione S-Transferase-Glutathione Interaction. J. Biomol. Screen, 21 (6). S. 596 - 608. THOUSAND OAKS: SAGE PUBLICATIONS INC. ISSN 1552-454X

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Abstract

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/Ni2+-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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Brenke, Jara K. UNSPECIFIED UNSPECIFIED UNSPECIFIED Salmina, Elena S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Ringelstetter, Larissa UNSPECIFIED UNSPECIFIED UNSPECIFIED Dornauer, Scarlett UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuzikov, Maria UNSPECIFIED UNSPECIFIED UNSPECIFIED Rothenaigner, Ina UNSPECIFIED UNSPECIFIED UNSPECIFIED Schorpp, Kenji UNSPECIFIED UNSPECIFIED UNSPECIFIED Giehler, Fabian UNSPECIFIED UNSPECIFIED UNSPECIFIED Gopalakrishnan, Jay UNSPECIFIED UNSPECIFIED UNSPECIFIED Kieser, Arnd UNSPECIFIED orcid.org/0000-0003-0783-1950 UNSPECIFIED Gul, Sheraz UNSPECIFIED UNSPECIFIED UNSPECIFIED Tetko, Igor V. UNSPECIFIED orcid.org/0000-0002-6855-0012 UNSPECIFIED Hadian, Kamyar UNSPECIFIED orcid.org/0000-0001-8727-2575 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-271556
DOI:	10.1177/1087057116639992
Journal or Publication Title:	J. Biomol. Screen
Volume:	21
Number:	6
Page Range:	S. 596 - 608
Date:	2016
Publisher:	SAGE PUBLICATIONS INC
Place of Publication:	THOUSAND OAKS
ISSN:	1552-454X
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language CENTER-DOT-S; CRYSTAL-STRUCTURES; FUSION PROTEINS; DRUG DISCOVERY; PURIFICATION; INHIBITORS; EVOLUTION; CHEMICALS; DATABASE; FILTERS Multiple languages Biochemical Research Methods; Biotechnology & Applied Microbiology; Chemistry, Analytical Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/27155