Parkinson's disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity
- Andrew B. West*,†,‡,
- Darren J. Moore*,†,‡,
- Saskia Biskup*,†,
- Artem Bugayenko*,†,
- Wanli W. Smith§,
- Christopher A. Ross†,§,¶,
- Valina L. Dawson*,†,¶,∥, and
- Ted M. Dawson*,†,¶,**
- *Institute for Cell Engineering, Departments of †Neurology, ¶Neuroscience, and ∥Physiology, and §Department of Psychiatry and Division of Neurobiology, 733 North Broadway, The Johns Hopkins University School of Medicine, Baltimore, MD 21205
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Edited by Solomon H. Snyder, The Johns Hopkins University School of Medicine, Baltimore, MD, and approved September 22, 2005 (received for review August 23, 2005)
Abstract
Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) cause late-onset Parkinson's disease (PD) with a clinical appearance indistinguishable from idiopathic PD. Initial studies suggest that LRRK2 mutations are the most common yet identified determinant of PD susceptibility, transmitted in an autosomal-dominant mode of inheritance. Herein, we characterize the LRRK2 gene and transcript in human brain and subclone the predominant ORF. Exogenously expressed LRRK2 protein migrates at ≈280 kDa and is present largely in the cytoplasm but also associates with the mitochondrial outer membrane. Familial-linked mutations G2019S or R1441C do not have an obvious effect on protein steady-state levels, turnover, or localization. However, in vitro kinase assays using full-length recombinant LRRK2 reveal an increase in activity caused by familial-linked mutations in both autophosphorylation and the phosphorylation of a generic substrate. These results suggest a gain-of-function mechanism for LRRK2-linked disease with a central role for kinase activity in the development of PD.
Footnotes
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↵ ** To whom correspondence should be addressed at: 733 North Broadway, Suite 731, Broadway Research Building, Baltimore, MD 21205. E-mail: tdawson{at}jhmi.edu.
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↵ ‡ A.B.W. and D.J.M. contributed equally to this work
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Author contributions: A.B.W., D.J.M., and T.M.D. designed research; A.B.W., D.J.M., and A.B. performed research; A.B.W. contributed new reagents/analytic tools; A.B.W., D.J.M., S.B., C.A.R., W.W.S., V.L.D., and T.M.D. analyzed data; and A.B.W., D.J.M., and T.M.D. wrote the paper.
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Conflict of interest statement: No conflicts declared.
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This paper was submitted directly (Track II) to the PNAS office.
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Abbreviations: HA, hemagglutinin; LRRK2, leucine-rich repeat kinase 2; MBP, myelin basic protein; MLK, mixed-lineage kinase; PD, Parkinson's disease.
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See Commentary on page 16535.
- Copyright © 2005, The National Academy of Sciences
