Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) are associated with familial and sporadic Parkinson's disease (PD). LRRK2 is a complex protein that consists of multiple domains, including predicted C-terminal WD40 repeats. In this study, we analyzed functional and molecular features conferred by the WD40 domain. Electron microscopic analysis of the purified LRRK2 C-terminal domain revealed doughnut-shaped particles, providing experimental evidence for its WD40 fold. We demonstrate that LRRK2 WD40 binds and sequesters synaptic vesicles via interaction with vesicle-associated proteins. In fact, a domain-based pulldown approach combined with mass spectrometric analysis identified LRRK2 as being part of a highly specific protein network involved in synaptic vesicle trafficking. In addition, we found that a C-terminal sequence variant associated with an increased risk of developing PD, G2385R, correlates with a reduced binding affinity of LRRK2 WD40 to synaptic vesicles. Our data demonstrate a critical role of the WD40 domain within LRRK2 function.
|Titolo:||Leucine-rich repeat kinase 2 binds to neuronal vesicles through : protein interactions mediated by its C-terminal WD40 domain|
|Parole Chiave:||Animals; Cells, Cultured; Humans; Mice; Mice, Inbred C57BL; Mutant Proteins; Mutation; Neurons; Neuropeptides; Neurotoxins; Parkinson Disease; Protein Binding; Protein Interaction Mapping; Protein-Serine-Threonine Kinases; Structure-Activity Relationship; Synapses; Synaptic Vesicles; Protein Interaction Domains and Motifs; Molecular Biology; Cell Biology|
|Settore Scientifico Disciplinare:||Settore BIO/14 - Farmacologia|
|Data di pubblicazione:||giu-2014|
|Digital Object Identifier (DOI):||10.1128/MCB.00914-13|
|Appare nelle tipologie:||01 - Articolo su periodico|