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Crystal structure of the WD40 domain dimer of LRRK2

Leucine-rich repeat kinase 2 (LRRK2) is a large multidomain protein with both a Ras of complex (ROC) domain and a kinase domain (KD) and, therefore, exhibits both GTPase and kinase activities. Human genetics studies have linked LRRK2 as a major genetic contributor to familial and sporadic Parkinson’...

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Autores principales: Zhang, Pengfei, Fan, Ying, Ru, Heng, Wang, Li, Magupalli, Venkat Giri, Taylor, Susan S., Alessi, Dario R., Wu, Hao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6358694/
https://www.ncbi.nlm.nih.gov/pubmed/30635421
http://dx.doi.org/10.1073/pnas.1817889116
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author Zhang, Pengfei
Fan, Ying
Ru, Heng
Wang, Li
Magupalli, Venkat Giri
Taylor, Susan S.
Alessi, Dario R.
Wu, Hao
author_facet Zhang, Pengfei
Fan, Ying
Ru, Heng
Wang, Li
Magupalli, Venkat Giri
Taylor, Susan S.
Alessi, Dario R.
Wu, Hao
author_sort Zhang, Pengfei
collection PubMed
description Leucine-rich repeat kinase 2 (LRRK2) is a large multidomain protein with both a Ras of complex (ROC) domain and a kinase domain (KD) and, therefore, exhibits both GTPase and kinase activities. Human genetics studies have linked LRRK2 as a major genetic contributor to familial and sporadic Parkinson’s disease (PD), a neurodegenerative movement disorder that inflicts millions worldwide. The C-terminal region of LRRK2 is a Trp-Asp-40 (WD40) domain with poorly defined biological functions but has been implicated in microtubule interaction. Here, we present the crystal structure of the WD40 domain of human LRRK2 at 2.6-Å resolution, which reveals a seven-bladed WD40 fold. The structure displays a dimeric assembly in the crystal, which we further confirm by measurements in solution. We find that structure-based and PD-associated disease mutations in the WD40 domain including the common G2385R polymorphism mainly compromise dimer formation. Assessment of full-length LRRK2 kinase activity by measuring phosphorylation of Rab10, a member of the family of Rab GTPases known to be important kinase substrates of LRRK2, shows enhancement of kinase activity by several dimerization-defective mutants including G2385R, although dimerization impairment does not always result in kinase activation. Furthermore, mapping of phylogenetically conserved residues onto the WD40 domain structure reveals surface patches that may be important for additional functions of LRRK2. Collectively, our analyses provide insights for understanding the structures and functions of LRRK2 and suggest the potential utility of LRRK2 kinase inhibitors in treating PD patients with WD40 domain mutations.
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spelling pubmed-63586942019-02-05 Crystal structure of the WD40 domain dimer of LRRK2 Zhang, Pengfei Fan, Ying Ru, Heng Wang, Li Magupalli, Venkat Giri Taylor, Susan S. Alessi, Dario R. Wu, Hao Proc Natl Acad Sci U S A Biological Sciences Leucine-rich repeat kinase 2 (LRRK2) is a large multidomain protein with both a Ras of complex (ROC) domain and a kinase domain (KD) and, therefore, exhibits both GTPase and kinase activities. Human genetics studies have linked LRRK2 as a major genetic contributor to familial and sporadic Parkinson’s disease (PD), a neurodegenerative movement disorder that inflicts millions worldwide. The C-terminal region of LRRK2 is a Trp-Asp-40 (WD40) domain with poorly defined biological functions but has been implicated in microtubule interaction. Here, we present the crystal structure of the WD40 domain of human LRRK2 at 2.6-Å resolution, which reveals a seven-bladed WD40 fold. The structure displays a dimeric assembly in the crystal, which we further confirm by measurements in solution. We find that structure-based and PD-associated disease mutations in the WD40 domain including the common G2385R polymorphism mainly compromise dimer formation. Assessment of full-length LRRK2 kinase activity by measuring phosphorylation of Rab10, a member of the family of Rab GTPases known to be important kinase substrates of LRRK2, shows enhancement of kinase activity by several dimerization-defective mutants including G2385R, although dimerization impairment does not always result in kinase activation. Furthermore, mapping of phylogenetically conserved residues onto the WD40 domain structure reveals surface patches that may be important for additional functions of LRRK2. Collectively, our analyses provide insights for understanding the structures and functions of LRRK2 and suggest the potential utility of LRRK2 kinase inhibitors in treating PD patients with WD40 domain mutations. National Academy of Sciences 2019-01-29 2019-01-11 /pmc/articles/PMC6358694/ /pubmed/30635421 http://dx.doi.org/10.1073/pnas.1817889116 Text en Copyright © 2019 the Author(s). Published by PNAS. http://creativecommons.org/licenses/by/4.0/ This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) .
spellingShingle Biological Sciences
Zhang, Pengfei
Fan, Ying
Ru, Heng
Wang, Li
Magupalli, Venkat Giri
Taylor, Susan S.
Alessi, Dario R.
Wu, Hao
Crystal structure of the WD40 domain dimer of LRRK2
title Crystal structure of the WD40 domain dimer of LRRK2
title_full Crystal structure of the WD40 domain dimer of LRRK2
title_fullStr Crystal structure of the WD40 domain dimer of LRRK2
title_full_unstemmed Crystal structure of the WD40 domain dimer of LRRK2
title_short Crystal structure of the WD40 domain dimer of LRRK2
title_sort crystal structure of the wd40 domain dimer of lrrk2
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6358694/
https://www.ncbi.nlm.nih.gov/pubmed/30635421
http://dx.doi.org/10.1073/pnas.1817889116
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