Leucine-rich repeat kinase 2 binds to neuronal vesicles through protein interactions mediated by its C-terminal WD40 domain

Mol Cell Biol. 2014 Jun;34(12):2147-61. doi: 10.1128/MCB.00914-13.

Giovanni Piccoli ¹, Franco Onofri ², Maria Daniela Cirnaru ³, Christoph J O Kaiser ⁴, Pravinkumar Jagtap ⁵, Andreas Kastenmüller ⁴, Francesca Pischedda ³, Antonella Marte ², Felix von Zweydorf ⁶, Andreas Vogt ⁷, Florian Giesert ⁸, Lifeng Pan ⁹, Flavia Antonucci ¹⁰, Christina Kiel ¹¹, Mingjie Zhang ⁹, Sevil Weinkauf ⁴, Michael Sattler ⁵, Carlo Sala ³, Michela Matteoli ¹⁰, Marius Ueffing ⁷, Christian Johannes Gloeckner ¹²

Affiliations

1 Helmholtz Zentrum München, German Research Center for Environmental Health, Research Unit Protein Science, Neuherberg, Germany Institute of Neuroscience, National Research Council, Milan, Italy.
2 Department of Experimental Medicine, University of Genoa, Genoa, Italy.
3 Institute of Neuroscience, National Research Council, Milan, Italy.
4 Center for Integrated Protein Science Munich and Technische Universität München, Department of Chemistry, Garching, Germany.
5 Center for Integrated Protein Science Munich and Technische Universität München, Department of Chemistry, Garching, Germany Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Structural Biology, Neuherberg, Germany.
6 Eberhard Karls University Tübingen, Institute for Ophthalmic Research, Medical Proteome Center, Tübingen, Germany.
7 Helmholtz Zentrum München, German Research Center for Environmental Health, Research Unit Protein Science, Neuherberg, Germany Eberhard Karls University Tübingen, Institute for Ophthalmic Research, Medical Proteome Center, Tübingen, Germany.
8 Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Developmental Genetics, Neuherberg, Germany.
9 Division of Life Science, Centre of Systems Biology and Human Health, Institute for Advanced Study and School of Science, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
10 Department of Biotechnology and Translational Medicine, University of Milan and Humanitas Clinical and Research Center, Rozzano, Italy.
11 CRG-EMBL System Biology Program, Centre de Regulació Genòmica UPF, Barcelona, Spain.
12 Helmholtz Zentrum München, German Research Center for Environmental Health, Research Unit Protein Science, Neuherberg, Germany Eberhard Karls University Tübingen, Institute for Ophthalmic Research, Medical Proteome Center, Tübingen, Germany [email protected].

PMID: 24687852 DOI: 10.1128/MCB.00914-13

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.

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