Leucine-Rich Repeat Kinase 2 Binds to Neuronal Vesicles through Protein Interactions Mediated by Its C-Terminal WD40 Domain

Giovanni Piccoli, Franco Onofri, Maria Daniela Cirnaru, Christoph J O Kaiser, Pravinkumar Jagtap, Andreas Kastenmüller, Francesca Pischedda, Antonella Marte, Felix von Zweydorf, Andreas Vogt, Florian Giesert, Lifeng Pan, Flavia Antonucci, Christina Kiel, Mingjie Zhang, Sevil Weinkauf, Michael Sattler, Carlo Sala, Michela Matteoli, Marius UeffingChristian Johannes Gloeckner

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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.

Original languageEnglish
Pages (from-to)2147-2161
Number of pages15
JournalMolecular and Cellular Biology
Volume34
Issue number12
DOIs
Publication statusPublished - Jun 1 2014

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Piccoli, G., Onofri, F., Cirnaru, M. D., Kaiser, C. J. O., Jagtap, P., Kastenmüller, A., Pischedda, F., Marte, A., von Zweydorf, F., Vogt, A., Giesert, F., Pan, L., Antonucci, F., Kiel, C., Zhang, M., Weinkauf, S., Sattler, M., Sala, C., Matteoli, M., ... Gloeckner, C. J. (2014). Leucine-Rich Repeat Kinase 2 Binds to Neuronal Vesicles through Protein Interactions Mediated by Its C-Terminal WD40 Domain. Molecular and Cellular Biology, 34(12), 2147-2161. https://doi.org/10.1128/MCB.00914-13