Format

Send to

Choose Destination
Mol Neurodegener. 2016 Jan 13;11:1. doi: 10.1186/s13024-015-0066-z.

LRRK2 phosphorylates pre-synaptic N-ethylmaleimide sensitive fusion (NSF) protein enhancing its ATPase activity and SNARE complex disassembling rate.

Author information

1
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. elisa.belluzzi@gmail.com.
2
Present Address: Rheumatology Unit, Department of Medicine - DIMED, University Hospital of Padova, Padova, Italy. elisa.belluzzi@gmail.com.
3
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. adriano.gonnelli@bio.unipd.it.
4
San Raffaele Scientific Park & University Vita-Salute San Raffaele, Milan, Italy. Cirnaru.mariadaniela@hsr.it.
5
Department of Experimental Medicine, University of Genova, Genova, Italy. antonella.marte@unige.it.
6
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. n.plotegher@ucl.ac.uk.
7
Present Address: Department of Cell and Developmental Biology, University College London, London, WC1E 6BT, UK. n.plotegher@ucl.ac.uk.
8
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. isabella.russo@unipd.it.
9
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. laura.civiero@unipd.it.
10
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. susanna.cogo@studenti.unipd.it.
11
San Raffaele Scientific Park & University Vita-Salute San Raffaele, Milan, Italy. mdolores.perez.carrion@gmail.com.
12
Department of Biomedical Sciences, University of Padova, Padova, Italy. cinzia.franchin@unipd.it.
13
Proteomics Center of Padova University, Padova, Italy. cinzia.franchin@unipd.it.
14
Department of Biomedical Sciences, University of Padova, Padova, Italy. giorgio.arrigoni@unipd.it.
15
Proteomics Center of Padova University, Padova, Italy. giorgio.arrigoni@unipd.it.
16
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. mariano.beltramini@unipd.it.
17
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. luigi.bubacco@unipd.it.
18
Department of Experimental Medicine, University of Genova, Genova, Italy. franco.onofri@unige.it.
19
San Raffaele Scientific Park & University Vita-Salute San Raffaele, Milan, Italy. piccoli.giovanni@hsr.it.
20
IN-CNR Milano, Milano, Italy. piccoli.giovanni@hsr.it.
21
Department of Biology, University of Padova, via Ugo Bassi 58/B, 35131, Padova, Italy. elisa.greggio@unipd.it.

Abstract

BACKGROUND:

Lrrk2, a gene linked to Parkinson's disease, encodes a large scaffolding protein with kinase and GTPase activities implicated in vesicle and cytoskeletal-related processes. At the presynaptic site, LRRK2 associates with synaptic vesicles through interaction with a panel of presynaptic proteins.

RESULTS:

Here, we show that LRRK2 kinase activity influences the dynamics of synaptic vesicle fusion. We therefore investigated whether LRRK2 phosphorylates component(s) of the exo/endocytosis machinery. We have previously observed that LRRK2 interacts with NSF, a hexameric AAA+ ATPase that couples ATP hydrolysis to the disassembling of SNARE proteins allowing them to enter another fusion cycle during synaptic exocytosis. Here, we demonstrate that NSF is a substrate of LRRK2 kinase activity. LRRK2 phosphorylates full-length NSF at threonine 645 in the ATP binding pocket of D2 domain. Functionally, NSF phosphorylated by LRRK2 displays enhanced ATPase activity and increased rate of SNARE complex disassembling. Substitution of threonine 645 with alanine abrogates LRRK2-mediated increased ATPase activity.

CONCLUSIONS:

Given that the most common Parkinson's disease LRRK2 G2019S mutation displays increased kinase activity, our results suggest that mutant LRRK2 may impair synaptic vesicle dynamics via aberrant phosphorylation of NSF.

PMID:
26758690
PMCID:
PMC4711005
DOI:
10.1186/s13024-015-0066-z
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for BioMed Central Icon for PubMed Central
Loading ...
Support Center