Martin, H.L., Turner, A.L., Higgins, J. et al. (14 more authors) (2023) Affimer-mediated locking of p21-activated kinase 5 in an intermediate activation state results in kinase inhibition. Cell Reports, 42 (10). 113184. ISSN 2211-1247
Abstract
Kinases are important therapeutic targets, and their inhibitors are classified according to their mechanism of action, which range from blocking ATP binding to covalent inhibition. Here, a mechanism of inhibition is highlighted by capturing p21-activated kinase 5 (PAK5) in an intermediate state of activation using an Affimer reagent that binds in the P+1 pocket. PAK5 was identified from a non-hypothesis-driven high-content imaging RNAi screen in urothelial cancer cells. Silencing of PAK5 resulted in reduced cell number, G1/S arrest, and enlargement of cells, suggesting it to be important in urothelial cancer cell line survival and proliferation. Affimer reagents were isolated to identify mechanisms of inhibition. The Affimer PAK5-Af17 recapitulated the phenotype seen with siRNA. Co-crystallization revealed that PAK5-Af17 bound in the P+1 pocket of PAK5, locking the kinase into a partial activation state. This mechanism of inhibition indicates that another class of kinase inhibitors is possible.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2023 The Authors. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | Affimer, high-content screening, urothelial cancer, PAK5, kinase, structural biology, signaling |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) |
Funding Information: | Funder Grant number Wellcome Trust 102174/B/13/Z |
Depositing User: | Symplectic Publications |
Date Deposited: | 19 Sep 2023 09:25 |
Last Modified: | 06 Oct 2023 14:29 |
Published Version: | https://www.cell.com/cell-reports/fulltext/S2211-1... |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.celrep.2023.113184 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:203432 |