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.
Leucine-rich repeat kinase 2 binds to neuronal vesicles through : protein interactions mediated by its C-terminal WD40 domain / G. Piccoli, F. Onofri, M.D. Cirnaru, C.J.O. Kaiser, P. Jagtap, A. Kastenmüller, F. Pischedda, A. Marte, F. von Zweydorf, A. Vogt, F. Giesert, L. Pan, F. Antonucci, C. Kiel, M. Zhang, S. Weinkauf, M. Sattler, C. Sala, M. Matteoli, M. Ueffing, C.J. Gloeckner. - In: MOLECULAR AND CELLULAR BIOLOGY. - ISSN 0270-7306. - 34:12(2014 Jun), pp. 2147-2161. [10.1128/MCB.00914-13]
Leucine-rich repeat kinase 2 binds to neuronal vesicles through : protein interactions mediated by its C-terminal WD40 domain
G. Piccoli;M.D. Cirnaru;F. Pischedda;F. Antonucci;M. Matteoli;
2014
Abstract
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.File | Dimensione | Formato | |
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