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April 19th, 2025
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Goethe-University Frankfurt
biochemistry
biorxiv

Covalent targeting leads to the development of LIMK1 isoform-selective inhibitors

Mandel, S.Open in Google Scholar•Hanke, T.Open in Google Scholar•Prendiville, N.Open in Google Scholar•Baena-Nuevo, M.Open in Google Scholar•Berger, L. M.Open in Google Scholar•Farges, F.Open in Google Scholar•Schwalm, M. P.Open in Google Scholar•Berger, B. T.Open in Google Scholar•Kraemer, A.Open in Google Scholar•Elson, L.Open in Google Scholaret al.

Selectivity for closely related isoforms of protein kinases is a major challenge in the design of drugs and chemical probes. Covalent targeting of unique cysteines is a potential strategy to achieve selectivity for highly conserved binding sites. Here, we used a pan-LIMK inhibitor to selectively probe LIMK1 over LIMK2 by targeting the LIMK1-specific cysteine C349 located in the glycine-rich loop region. Binding kinetics of both non-covalent and covalent LIMK inhibitors were investigated, and the fast on-rate and small size of type-I inhibitors were used in the design of a covalent LIMK1 inhibitor. The developed cell-active, isoform-selective LIMK1 inhibitor showed excellent proteome-wide selectivity in pull-down assays, enabling studies of LIMK1 isoform-selective functions in cellular model systems and providing a versatile chemical tool for studies of the LIMK signalling pathway.

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