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LIM domain kinase 1 (LIMK1) is a key regulator of actin dynamics. It is thereby a potential therapeutic target for the prevention of fragile X syndrome and amyotrophic lateral sclerosis. Herein, we use X-ray crystallography and activity assays to describe how LIMK1 accomplishes substrate specificity, to suggest a unique 'rock-and-poke' mechanism of catalysis and to explore the regulation of the kinase by activation loop phosphorylation. Based on these findings, a differential scanning fluorimetry assay and a RapidFire mass spectrometry activity assay were established, leading to the discovery and confirmation of a set of small-molecule LIMK1 inhibitors. Interestingly, several of the inhibitors were inactive towards the closely related isoform LIMK2. Finally, crystal structures of the LIMK1 kinase domain in complex with inhibitors (PF-477736 and staurosporine, respectively) are presented, providing insights into LIMK1 plasticity upon inhibitor binding.

Original publication

DOI

10.1042/BCJ20190517

Type

Journal article

Journal

Biochem J

Publication Date

15/11/2019

Volume

476

Pages

3197 - 3209

Keywords

LIMK1, kinase, reaction mechanism, small-molecule inhibitors, substrate recognition, Catalysis, Crystallography, Drug Design, Humans, Lim Kinases, Models, Molecular, Phosphorylation, Protein Kinase Inhibitors, Substrate Specificity