Identification of Small-Molecule Frequent Hitters of Glutathione S-Transferase-Glutathione Interaction
| Autor: | Kamyar Hadian, Maria Kuzikov, Kenji Schorpp, Fabian Giehler, Elena Salmina, Igor V. Tetko, Jay Gopalakrishnan, Sheraz Gul, Jara Kerstin Brenke, Ina Rothenaigner, Larissa Ringelstetter, Arnd Kieser, Scarlett Dornauer |
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| Přispěvatelé: | Publica |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
High-throughput screening 01 natural sciences Biochemistry Substrate Specificity Analytical Chemistry Small Molecule Libraries 03 medical and health sciences chemistry.chemical_compound High-Throughput Screening Assays Humans Protein Interaction Maps Glutathione Transferase biology Glutathione Small molecule Molecular biology Enzyme assay 0104 chemical sciences 010404 medicinal & biomolecular chemistry 030104 developmental biology Glutathione S-transferase chemistry biology.protein Molecular Medicine Protein Interaction Map Biotechnology |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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