title: Alterations in the mitochondrial proteome of neuroblastoma cells in response to complex 1 inhibition.
abstract: Increasing evidence points to mitochondrial dysfunction in Parkinson's disease (PD) associated with complex I dysfunction, but the exact pathways which lead to cell death have not been resolved. 2D-gel electrophoresis profiles of isolated mitochondria from neuroblastoma cells treated with subcytotoxic concentrations of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a well-characterized complex I inhibitor, were assessed to identify associated targets. Up to 27 differentially expressed proteins were observed, of which 16 were identified using peptide mass fingerprinting. Changes in protein levels were validated by immunoprobing 1D blots, confirming increases in heat shock cognate 71 kDa (Hsc70), 60 kDa heat shock protein (Hsp60), fumarase, glutamate oxaloacetate transaminase 2, ATP synthase subunit d, and voltage-dependent anion-channel 1 (VDAC1). Immunoprobing of 2D blots revealed isoform changes in Hsc70, Hsp60, and VDAC1. Subcytotoxic concentrations of MPTP modulated a host of mitochondrial proteins including chaperones, metabolic enzymes, oxidative phosphorylation-related proteins, an inner mitochondrial protein (mitofilin), and an outer mitochondrial membrane protein (VDAC1). Early changes in chaperones suggest a regulated link between complex 1 inhibition and protein folding. VDAC1, a multifunctional protein, may have a key role in signaling between mitochondria and the rest of the cell prior to cell death. Our work provides new important information of relevance to PD.
authors.affiliation: School of Science and Technology, Nottingham Trent University, Clifton Lane, NG11 8NS Nottingham, UK.
authors.forename: Florence
authors.initials: F
authors.lastname: Burté
authors.forename: Luigi A
authors.initials: LA
authors.lastname: De Girolamo
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Hargreaves
authors.forename: E Ellen
authors.initials: EE
authors.lastname: Billett
mesh_terms: D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000494:pharmacology; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D045744:Cell Line, Tumor; D042967:Electron Transport Complex I / Q000037:antagonists & inhibitors* / Q000187:drug effects; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008928:Mitochondria / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000032:analysis*; D009447:Neuroblastoma / Q000737:chemistry*; D009498:Neurotoxins / Q000633:toxicity; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods
chemical_list: D015415:Biomarkers; D024101:Mitochondrial Proteins; D009498:Neurotoxins; D020543:Proteome; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D042967:Electron Transport Complex I