Exploring the Impact of PARK2 Mutations on the Total and Mitochondrial Proteome of Human Skin Fibroblasts.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:32596240", "kind": "science-record", "revision": 7, "content_hash": "sha256:69fd9e842381f471ca30f0655249edcbdcbaaa0e1574289fb69628fa09e057b2", "created_at": "2026-11-01T15:41:04Z", "updated_at": "2026-11-01T22:01:49Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Exploring the Impact of PARK2 Mutations on the Total and Mitochondrial Proteome of Human Skin Fibroblasts.", "doi": "10.3389/fcell.2020.00423", "pmid": "32596240", "authors": ["Mara Zilocchi", "Ilaria Colugnat", "Marta Lualdi", "Monica Meduri", "Federica Marini", "Victor Corasolla Carregari", "Mohamed Taha Moutaoufik", "Sadhna Phanse", "Luisa Pieroni", "Mohan Babu", "Barbara Garavaglia", "Mauro Fasano", "Tiziana Alberio"], "journal": "Frontiers in cell and developmental biology", "year": 2020, "abstract": "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.", "claims": [{"id": "claim-data-access", "statement": "The datasets generated for this study can be found in the ProteomeXchange (Accession: PXD015880).", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:32596240", "locator": "sections[23] \"Data Availability Statement\" paragraph 2", "quote": "The datasets generated for this study can be found in the ProteomeXchange (Accession: PXD015880)."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": null, "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:32596240"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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On t"], "order_basis": "paper_text_order:919", "id": "assay:2"}, {"role": "data_generation", "definition_id": "cell_culture", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["tures and Treatments Primary skin fibroblast cell lines were cultured in high glucose Dulbecco’s modified Eagle’s", "ents Primary skin fibroblast cell lines were cultured in high glucose Dulbecco’s modified Eagle’s med"], "order_basis": "paper_text_order:2276", "id": "assay:3"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["utated patients and five control subjects by western blot. As shown in Figure 1, a complete loss of Pa", "n PARK2-mutated patients. (A) Representative western blot image of Parkin protein in primary skin fibr", "and optic atrophy 1 (OPA1), were measured by western blot analysis (Figures 4C,E). As shown in Figures", "s mean ± SEM. ∗p = 0.026. (C) Representative western blot image of MFN1 protein in primary skin fibrob"], "order_basis": "paper_text_order:3354", "id": "assay:4"}, {"role": "data_generation", "definition_id": "cell_imaging", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["visualize the mitochondrial network through confocal microscopy, in the absence and in the presence of CCCP", "e the mitochondrial network through confocal microscopy, in the absence and in the presence of CCCP"], "order_basis": "paper_text_order:4391", "id": "assay:5"}, {"role": "data_generation", "definition_id": "ihc", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["in PARK2-mutated samples. (A) Representative immunofluorescence images of primary skin fibroblasts from one", "of Rab7A to mitochondria was investigated by immunofluorescence. To this end, quantification analysis of Rab"], "order_basis": "paper_text_order:5515", "id": "assay:6"}, {"role": "analysis", "definition_id": "group_comparison", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 0.8, "evidence": ["Wilcoxon"], "order_basis": "consumes_earlier_output", "id": "assay:7"}]}