TY - JOUR
T1 - Exploring the Impact of PARK2 Mutations on the Total and Mitochondrial Proteome of Human Skin Fibroblasts
AU - Zilocchi, Mara
AU - Colugnat, Ilaria
AU - Lualdi, Marta
AU - Meduri, Monica
AU - Marini, Federica
AU - Corasolla Carregari, Victor
AU - Moutaoufik, Mohamed Taha
AU - Phanse, Sadhna
AU - Pieroni, Luisa
AU - Babu, Mohan
AU - Garavaglia, Barbara
AU - Fasano, Mauro
AU - Alberio, Tiziana
N1 - Funding Information:
We acknowledge the Telethon Network of Genetic Biobanks for providing us the samples of PD and control subjects and all the donors. Funding. This work was supported by grants from the Canadian Institutes of Health Research (MOP-125952; FDN-154318), as well as the Parkinson Society of Canada (2014-673) and National Institutes of Health (R01GM106019) to MB, and from Teofilo Rossi di Montelera Foundation to MF.
Publisher Copyright:
© Copyright © 2020 Zilocchi, Colugnat, Lualdi, Meduri, Marini, Corasolla Carregari, Moutaoufik, Phanse, Pieroni, Babu, Garavaglia, Fasano and Alberio.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/6/11
Y1 - 2020/6/11
N2 - Mutations in PARK2 gene are the most frequent cause of familial forms of Parkinson’s disease (PD). This gene encodes Parkin, an E3 ubiquitin ligase involved in several cellular mechanisms, including mitophagy. Parkin loss-of-function is responsible for the cellular accumulation of damaged mitochondria, which in turn determines an increment of reactive oxygen species (ROS) levels, lower ATP production, and apoptosis activation. Given the importance of mitochondrial dysfunction and mitophagy impairment in PD pathogenesis, the aim of the present study was to investigate both total and mitochondrial proteome alterations in human skin fibroblasts of PARK2-mutated patients. To this end, both total and mitochondria-enriched protein fractions from fibroblasts of five PARK2-mutated patients and five control subjects were analyzed by quantitative shotgun proteomics to identify proteins specifically altered by Parkin mutations (mass spectrometry proteomics data have been submitted to ProteomeXchange with the identifier PXD015880). Both the network-based and gene set enrichment analyses pointed out pathways in which Rab GTPase proteins are involved. To have a more comprehensive view of the mitochondrial alterations due to PARK2 mutations, we investigated the impact of Parkin loss on mitochondrial function and network morphology. We unveiled that the mitochondrial membrane potential was reduced in PARK2-mutated patients, without inducing PINK1 accumulation, even when triggered with the ionophore carbonyl cyanide m-chlorophenylhydrazone (CCCP). Lastly, the analysis of the mitochondrial network morphology did not reveal any significant alterations in PARK2-mutated patients compared to control subjects. Thus, our results suggested that the network morphology was not influenced by the mitochondrial depolarization and by the lack of Parkin, revealing a possible impairment of fission and, more in general, of mitochondrial dynamics. In conclusion, the present work highlighted new molecular factors and pathways altered by PARK2 mutations, which will unravel possible biochemical pathways altered in the sporadic form of PD.
AB - Mutations in PARK2 gene are the most frequent cause of familial forms of Parkinson’s disease (PD). This gene encodes Parkin, an E3 ubiquitin ligase involved in several cellular mechanisms, including mitophagy. Parkin loss-of-function is responsible for the cellular accumulation of damaged mitochondria, which in turn determines an increment of reactive oxygen species (ROS) levels, lower ATP production, and apoptosis activation. Given the importance of mitochondrial dysfunction and mitophagy impairment in PD pathogenesis, the aim of the present study was to investigate both total and mitochondrial proteome alterations in human skin fibroblasts of PARK2-mutated patients. To this end, both total and mitochondria-enriched protein fractions from fibroblasts of five PARK2-mutated patients and five control subjects were analyzed by quantitative shotgun proteomics to identify proteins specifically altered by Parkin mutations (mass spectrometry proteomics data have been submitted to ProteomeXchange with the identifier PXD015880). Both the network-based and gene set enrichment analyses pointed out pathways in which Rab GTPase proteins are involved. To have a more comprehensive view of the mitochondrial alterations due to PARK2 mutations, we investigated the impact of Parkin loss on mitochondrial function and network morphology. We unveiled that the mitochondrial membrane potential was reduced in PARK2-mutated patients, without inducing PINK1 accumulation, even when triggered with the ionophore carbonyl cyanide m-chlorophenylhydrazone (CCCP). Lastly, the analysis of the mitochondrial network morphology did not reveal any significant alterations in PARK2-mutated patients compared to control subjects. Thus, our results suggested that the network morphology was not influenced by the mitochondrial depolarization and by the lack of Parkin, revealing a possible impairment of fission and, more in general, of mitochondrial dynamics. In conclusion, the present work highlighted new molecular factors and pathways altered by PARK2 mutations, which will unravel possible biochemical pathways altered in the sporadic form of PD.
KW - interactome
KW - mitochondria
KW - mitophagy
KW - Parkin (PARK2)
KW - Parkinson’s Disease
KW - proteomic alterations
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UR - http://www.scopus.com/inward/citedby.url?scp=85087024934&partnerID=8YFLogxK
U2 - 10.3389/fcell.2020.00423
DO - 10.3389/fcell.2020.00423
M3 - Article
AN - SCOPUS:85087024934
VL - 8
JO - Frontiers in Cell and Developmental Biology
JF - Frontiers in Cell and Developmental Biology
SN - 2296-634X
M1 - 423
ER -