Quantitative proteomic analysis of Parkin substrates in Drosophila neurons
dc.contributor.author | Martínez Zárate, Aitor | |
dc.contributor.author | Lectez, Benoit | |
dc.contributor.author | Ramírez Sánchez, Juan Manuel | |
dc.contributor.author | Popp, Oliver | |
dc.contributor.author | Sutherland, James D. | |
dc.contributor.author | Urbé, Sylvie | |
dc.contributor.author | Dittmar, Gunnar | |
dc.contributor.author | Clague, Michael J. | |
dc.contributor.author | Mayor Martínez, Ugo | |
dc.date.accessioned | 2018-06-13T12:23:36Z | |
dc.date.available | 2018-06-13T12:23:36Z | |
dc.date.issued | 2017-04-11 | |
dc.identifier.citation | Molecular Neurodegeneration 12 : (2017) // Article ID 29 | es_ES |
dc.identifier.issn | 1750-1326 | |
dc.identifier.uri | http://hdl.handle.net/10810/27512 | |
dc.description.abstract | Background: Parkin (PARK2) is an E3 ubiquitin ligase that is commonly mutated in Familial Parkinson's Disease (PD). In cell culture models, Parkin is recruited to acutely depolarised mitochondria by PINK1. PINK1 activates Parkin activity leading to ubiquitination of multiple proteins, which in turn promotes clearance of mitochondria by mitophagy. Many substrates have been identified using cell culture models in combination with depolarising drugs or proteasome inhibitors, but not in more physiological settings. Methods: Here we utilized the recently introduced BioUb strategy to isolate ubiquitinated proteins in flies. Following Parkin Wild-Type (WT) and Parkin Ligase dead (LD) expression we analysed by mass spectrometry and stringent bioinformatics analysis those proteins differentially ubiquitinated to provide the first survey of steady state Parkin substrates using an in vivo model. We further used an in vivo ubiquitination assay to validate one of those substrates in SH-SY5Y cells. Results: We identified 35 proteins that are more prominently ubiquitinated following Parkin over-expression. These include several mitochondrial proteins and a number of endosomal trafficking regulators such as v-ATPase sub-units, Syx5/STX5, ALiX/PDCD6IP and Vps4. We also identified the retromer component, Vps35, another PD-associated gene that has recently been shown to interact genetically with parkin. Importantly, we validated Parkin-dependent ubiquitination of VPS35 in human neuroblastoma cells. Conclusions: Collectively our results provide new leads to the possible physiological functions of Parkin activity that are not overtly biased by acute mitochondrial depolarisation. | es_ES |
dc.description.sponsorship | A.M. is a recipient of a CIC bioGUNE/Liverpool University studentship. U.M. is a recipient of a MINECO grant (SAF2013-44782-P) co-financed with FEDER funds. | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Biomed Central | es_ES |
dc.relation | info:eu-repo/grantAgreement/MINECO/SAF2013-44782-P | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.subject | Parkin (PARK2) | es_ES |
dc.subject | Parkin substrates | es_ES |
dc.subject | ubiquitination | es_ES |
dc.subject | VPS35 | es_ES |
dc.subject | neurodegeneration | es_ES |
dc.subject | in vivo | es_ES |
dc.subject | Drosophila melanogaster | es_ES |
dc.subject | Parkinson's disease (PD) | es_ES |
dc.subject | Alzheimer's disease (AD) | es_ES |
dc.subject | label free quantification (LFQ) | es_ES |
dc.subject | ubiquitin ligase activity | es_ES |
dc.subject | mitochondrial dysfunction | es_ES |
dc.subject | retromer complex | es_ES |
dc.subject | alpha-synuclein | es_ES |
dc.subject | pink1/parkin-mediated mitophagy | es_ES |
dc.subject | biotinylated ubiquitin | es_ES |
dc.subject | pathogenic mutations | es_ES |
dc.subject | endosomal pathway | es_ES |
dc.subject | activate Parkin | es_ES |
dc.title | Quantitative proteomic analysis of Parkin substrates in Drosophila neurons | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. | es_ES |
dc.rights.holder | Atribución 3.0 España | * |
dc.relation.publisherversion | https://molecularneurodegeneration.biomedcentral.com/articles/10.1186/s13024-017-0170-3 | es_ES |
dc.identifier.doi | 10.1186/s13024-017-0170-3 | |
dc.departamentoes | Bioquímica y biología molecular | es_ES |
dc.departamentoeu | Biokimika eta biologia molekularra | es_ES |
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