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Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97

Biosensor techniques have become increasingly important for fragment-based drug discovery during the last years. The AAA+ ATPase p97 is an essential protein with key roles in protein homeostasis and a possible target for cancer chemotherapy. Currently available p97 inhibitors address its ATPase acti...

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Autores principales: Bothe, Sebastian, Hänzelmann, Petra, Böhler, Stephan, Kehrein, Josef, Zehe, Markus, Wiedemann, Christoph, Hellmich, Ute A., Brenk, Ruth, Schindelin, Hermann, Sotriffer, Christoph
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9814400/
https://www.ncbi.nlm.nih.gov/pubmed/36697690
http://dx.doi.org/10.1038/s42004-022-00782-5
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author Bothe, Sebastian
Hänzelmann, Petra
Böhler, Stephan
Kehrein, Josef
Zehe, Markus
Wiedemann, Christoph
Hellmich, Ute A.
Brenk, Ruth
Schindelin, Hermann
Sotriffer, Christoph
author_facet Bothe, Sebastian
Hänzelmann, Petra
Böhler, Stephan
Kehrein, Josef
Zehe, Markus
Wiedemann, Christoph
Hellmich, Ute A.
Brenk, Ruth
Schindelin, Hermann
Sotriffer, Christoph
author_sort Bothe, Sebastian
collection PubMed
description Biosensor techniques have become increasingly important for fragment-based drug discovery during the last years. The AAA+ ATPase p97 is an essential protein with key roles in protein homeostasis and a possible target for cancer chemotherapy. Currently available p97 inhibitors address its ATPase activity and globally impair p97-mediated processes. In contrast, inhibition of cofactor binding to the N-domain by a protein-protein-interaction inhibitor would enable the selective targeting of specific p97 functions. Here, we describe a biolayer interferometry-based fragment screen targeting the N-domain of p97 and demonstrate that a region known as SHP-motif binding site can be targeted with small molecules. Guided by molecular dynamics simulations, the binding sites of selected screening hits were postulated and experimentally validated using protein- and ligand-based NMR techniques, as well as X-ray crystallography, ultimately resulting in the first structure of a small molecule in complex with the N-domain of p97. The identified fragments provide insights into how this region could be targeted and present first chemical starting points for the development of a protein-protein interaction inhibitor preventing the binding of selected cofactors to p97.
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spelling pubmed-98144002023-01-10 Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97 Bothe, Sebastian Hänzelmann, Petra Böhler, Stephan Kehrein, Josef Zehe, Markus Wiedemann, Christoph Hellmich, Ute A. Brenk, Ruth Schindelin, Hermann Sotriffer, Christoph Commun Chem Article Biosensor techniques have become increasingly important for fragment-based drug discovery during the last years. The AAA+ ATPase p97 is an essential protein with key roles in protein homeostasis and a possible target for cancer chemotherapy. Currently available p97 inhibitors address its ATPase activity and globally impair p97-mediated processes. In contrast, inhibition of cofactor binding to the N-domain by a protein-protein-interaction inhibitor would enable the selective targeting of specific p97 functions. Here, we describe a biolayer interferometry-based fragment screen targeting the N-domain of p97 and demonstrate that a region known as SHP-motif binding site can be targeted with small molecules. Guided by molecular dynamics simulations, the binding sites of selected screening hits were postulated and experimentally validated using protein- and ligand-based NMR techniques, as well as X-ray crystallography, ultimately resulting in the first structure of a small molecule in complex with the N-domain of p97. The identified fragments provide insights into how this region could be targeted and present first chemical starting points for the development of a protein-protein interaction inhibitor preventing the binding of selected cofactors to p97. Nature Publishing Group UK 2022-12-07 /pmc/articles/PMC9814400/ /pubmed/36697690 http://dx.doi.org/10.1038/s42004-022-00782-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Bothe, Sebastian
Hänzelmann, Petra
Böhler, Stephan
Kehrein, Josef
Zehe, Markus
Wiedemann, Christoph
Hellmich, Ute A.
Brenk, Ruth
Schindelin, Hermann
Sotriffer, Christoph
Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97
title Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97
title_full Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97
title_fullStr Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97
title_full_unstemmed Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97
title_short Fragment screening using biolayer interferometry reveals ligands targeting the SHP-motif binding site of the AAA+ ATPase p97
title_sort fragment screening using biolayer interferometry reveals ligands targeting the shp-motif binding site of the aaa+ atpase p97
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9814400/
https://www.ncbi.nlm.nih.gov/pubmed/36697690
http://dx.doi.org/10.1038/s42004-022-00782-5
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