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- Title
Lessons from LIMK1 enzymology and their impact on inhibitor design.
- Authors
Salah, Eidarus; Chatterjee, Deep; Beltrami, Alessandra; Tumber, Anthony; Preuss, Franziska; Canning, Peter; Chaikuad, Apirat; Knaus, Petra; Knapp, Stefan; Bullock, Alex N.; Mathea, Sebastian
- Abstract
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.
- Subjects
FRAGILE X syndrome; AMYOTROPHIC lateral sclerosis; ENZYMOLOGY; KINASE regulation; ACETOLACTATE synthase; MASS spectrometry; X-ray crystallography; GENE amplification
- Publication
Biochemical Journal, 2019, Vol 476, Issue 21, p3197
- ISSN
0264-6021
- Publication type
Article
- DOI
10.1042/BCJ20190517