Lessons from LIMK1 enzymology and their impact on inhibitor design
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...
| Main Authors: | , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Portland Press
2019
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| _version_ | 1826295485365747712 |
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| author | Salah, E Chatterjee, D Beltrami, A Tumber, A Preuss, F Canning, P Chaikuad, A Knaus, P Knapp, S Bullock, A Mathea, S |
| author_facet | Salah, E Chatterjee, D Beltrami, A Tumber, A Preuss, F Canning, P Chaikuad, A Knaus, P Knapp, S Bullock, A Mathea, S |
| author_sort | Salah, E |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-07T04:01:46Z |
| format | Journal article |
| id | oxford-uuid:c4cf49d7-fe81-4957-b132-2a8880579718 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T04:01:46Z |
| publishDate | 2019 |
| publisher | Portland Press |
| record_format | dspace |
| spelling | oxford-uuid:c4cf49d7-fe81-4957-b132-2a88805797182022-03-27T06:26:24ZLessons from LIMK1 enzymology and their impact on inhibitor designJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:c4cf49d7-fe81-4957-b132-2a8880579718EnglishSymplectic Elements at OxfordPortland Press2019Salah, EChatterjee, DBeltrami, ATumber, APreuss, FCanning, PChaikuad, AKnaus, PKnapp, SBullock, AMathea, SLIM 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. |
| spellingShingle | Salah, E Chatterjee, D Beltrami, A Tumber, A Preuss, F Canning, P Chaikuad, A Knaus, P Knapp, S Bullock, A Mathea, S Lessons from LIMK1 enzymology and their impact on inhibitor design |
| title | Lessons from LIMK1 enzymology and their impact on inhibitor design |
| title_full | Lessons from LIMK1 enzymology and their impact on inhibitor design |
| title_fullStr | Lessons from LIMK1 enzymology and their impact on inhibitor design |
| title_full_unstemmed | Lessons from LIMK1 enzymology and their impact on inhibitor design |
| title_short | Lessons from LIMK1 enzymology and their impact on inhibitor design |
| title_sort | lessons from limk1 enzymology and their impact on inhibitor design |
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