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‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement

Protein kinases catalyse the addition of phosphate groups to Ser/Thr and Tyr residues in cognate substrates and are mutated or hyperactive in a variety of diseases, making them important targets for rationally designed drugs. A good example is the Parkinson's disease-associated kinase, leucine-...

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Autor principal: Eyers, Patrick A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5095898/
https://www.ncbi.nlm.nih.gov/pubmed/27621483
http://dx.doi.org/10.1042/BCJ20160671C
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author_facet Eyers, Patrick A.
author_sort Eyers, Patrick A.
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description Protein kinases catalyse the addition of phosphate groups to Ser/Thr and Tyr residues in cognate substrates and are mutated or hyperactive in a variety of diseases, making them important targets for rationally designed drugs. A good example is the Parkinson's disease-associated kinase, leucine-rich repeat kinase 2 (LRRK2), which is mutated (and probably hyperactive) in a small, but significant, subset of patients. An exciting new approach for personalised therapy is the development of central nervous system (CNS)-active small-molecule kinase inhibitors, which could be employed to ‘normalise’ LRRK2 signalling in affected cell types. However, the development of such drugs requires validated assays for the analysis of target engagement and the assembly of a set of tools for interrogating LRRK2, and its substrates, both in vitro and in vivo. A new study published in the Biochemical Journal by Ito et al. establishes that a ‘Phos-tag’™-binding assay can be exploited to measure phosphorylation of a recently identified LRRK2 substrate (Ras-related protein in brain 10 (Rab10)), and to compare and contrast relative catalytic output from disease-associated LRRK2 mutants. Powerful in vivo chemical genetic approaches are also disclosed, in which the catalytic activity of LRRK2 is unequivocally linked to the extent of Rab10 phosphorylation and the effects of chemically distinct LRRK2 inhibitors are matched with on-target inhibition mechanisms mediated through LRRK2 and its substrate Rab10. These important findings should simplify the generic analysis of Rab10 phosphorylation in model biological systems and are likely to be applicable to other substrates of LRRK2 (or indeed other kinases) for which phospho-specific antibodies are either absent or unsatisfactory.
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spelling pubmed-50958982016-11-08 ‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement Eyers, Patrick A. Biochem J Commentaries Protein kinases catalyse the addition of phosphate groups to Ser/Thr and Tyr residues in cognate substrates and are mutated or hyperactive in a variety of diseases, making them important targets for rationally designed drugs. A good example is the Parkinson's disease-associated kinase, leucine-rich repeat kinase 2 (LRRK2), which is mutated (and probably hyperactive) in a small, but significant, subset of patients. An exciting new approach for personalised therapy is the development of central nervous system (CNS)-active small-molecule kinase inhibitors, which could be employed to ‘normalise’ LRRK2 signalling in affected cell types. However, the development of such drugs requires validated assays for the analysis of target engagement and the assembly of a set of tools for interrogating LRRK2, and its substrates, both in vitro and in vivo. A new study published in the Biochemical Journal by Ito et al. establishes that a ‘Phos-tag’™-binding assay can be exploited to measure phosphorylation of a recently identified LRRK2 substrate (Ras-related protein in brain 10 (Rab10)), and to compare and contrast relative catalytic output from disease-associated LRRK2 mutants. Powerful in vivo chemical genetic approaches are also disclosed, in which the catalytic activity of LRRK2 is unequivocally linked to the extent of Rab10 phosphorylation and the effects of chemically distinct LRRK2 inhibitors are matched with on-target inhibition mechanisms mediated through LRRK2 and its substrate Rab10. These important findings should simplify the generic analysis of Rab10 phosphorylation in model biological systems and are likely to be applicable to other substrates of LRRK2 (or indeed other kinases) for which phospho-specific antibodies are either absent or unsatisfactory. Portland Press Ltd. 2016-09-15 2016-09-12 /pmc/articles/PMC5095898/ /pubmed/27621483 http://dx.doi.org/10.1042/BCJ20160671C Text en © 2016 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0) .
spellingShingle Commentaries
Eyers, Patrick A.
‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement
title ‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement
title_full ‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement
title_fullStr ‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement
title_full_unstemmed ‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement
title_short ‘Up with the LRRK’: a phosphorylated Rab10 assay for evaluation of LRRK2 activity and inhibitor engagement
title_sort ‘up with the lrrk’: a phosphorylated rab10 assay for evaluation of lrrk2 activity and inhibitor engagement
topic Commentaries
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5095898/
https://www.ncbi.nlm.nih.gov/pubmed/27621483
http://dx.doi.org/10.1042/BCJ20160671C
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