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Mol. Cell. Proteomics 10:M110.001172 (2011)
Verlagsversion Forschungsdaten Forschungsdaten DOI
Mutations in human leucine-rich repeat kinase 2 (Lrrk2), a protein of yet unknown function, are linked to Parkinson's disease caused by degeneration of midbrain dopaminergic neurons. The protein comprises several domains including a GTPase- and a kinase domain both affected by several pathogenic mutations. To elucidate the molecular interaction network of endogenous Lrrk2 under stoichiometric constraints, we applied QUICK (quantitative immunoprecipitation combined with knockdown) in NIH3T3 cells. The identified interactome reveals actin isoforms as well as actin-associated proteins involved in actin filament assembly, organization, rearrangement and maintenance, suggesting that the biological function of Lrrk2 is linked to cytoskeletal dynamics. In fact, we demonstrate Lrrk2 de novo binding to F-actin and its ability to modulate its assembly in vitro. When tested in intact cells, knockdown of Lrrk2 causes morphological alterations in NIH3T3 cells. In developing dopaminergic midbrain primary neurons, Lrrk2 knockdown results in shortened neurite processes, indicating a physiological role of Lrrk2 in cytoskeletal organization and dynamics of dopaminergic neurons. Hence, our results demonstrate that molecular interactions as well as the physiological function of Lrrk2 are closely related to the organization of the actin-based cytoskeleton, a crucial feature of neuronal development and neuron function.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Mass spectrometry; Networks ; Neurodegenerative diseases; Protein-protein interactions; SILAC; siRNA; Leucine-rich repeat kinase 2 (Lrrk2); Actin cytoskeleton; Quantitative immunoprecipitation combined with knockdown (QUICK); Dopaminergic neurons; Parkinson’s disease; Neurite outgrowth
ISSN (print) / ISBN 1535-9476
Zeitschrift Molecular and Cellular Proteomics
Quellenangaben Band: 10, Heft: 1, Artikelnummer: M110.001172
Verlag American Society for Biochemistry and Molecular Biology
Begutachtungsstatus Peer reviewed
Institut(e) Core Facility Metabolomics & Proteomics (CF-MPC)