Lessons from LIMK1 enzymology and their impact on inhibitor design

DOI: 10.1042/BCJ20190517

Authors: Eidarus Salah (Target Discovery Institute; Structural Genomics Consortium, University of Oxford) , Deep Chatterjee (Structural Genomics Consortium, Goethe-University Frankfurt) , Alessandra Beltrami (Structural Genomics Consortium, University of Oxford) , Anthony Tumber (Target Discovery Institute; Structural Genomics Consortium, University of Oxford) , Franziska Preuss (Structural Genomics Consortium, Goethe-University Frankfurt) , Peter Canning (Structural Genomics Consortium, University of Oxford) , Apirat Chaikuad (Structural Genomics Consortium, Goethe-University Frankfurt) , Petra Knaus (Freie Universität Berlin) , Stefan Knapp (Structural Genomics Consortium, Goethe-University Frankfurt) , Alex N. Bullock (Structural Genomics Consortium, University of Oxford) , Sebastian Mathea (Structural Genomics Consortium, University of Oxford; Goethe-University Frankfurt)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Biochemical Journal

State: Published (Approved)
Published: October 2019
Diamond Proposal Number(s): 6391 , 10619

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.

Journal Keywords: Kinase; Small-molecule inhibitors; Reaction mechanism; Substrate recognition; LIMK1

Subject Areas: Biology and Bio-materials


Instruments: I02-Macromolecular Crystallography , I03-Macromolecular Crystallography , I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography

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