title: Subcomplexes of human ATP synthase mark mitochondrial biosynthesis disorders.
abstract: OBJECTIVE
  
  
  
  METHODS
  We describe biochemically and clinically relevant aspects of mitochondrial ATP synthase, the enzyme that supplies most ATP for the cells energy demand.
  
  
  RESULTS
  Analyzing human Rho zero cells we could identify three subcomplexes of ATP synthase: F1 catalytic domain, F1 domain with bound natural IF1 inhibitor protein, and F1-c subcomplex, an assembly of F1 domain and a ring of F(O)-subunits c. Large amounts of F1 subcomplexes accumulated also in mitochondria of patients with specific mitochondrial disorders. By quantifying the F1 subcomplexes and other oxidative phosphorylation complexes in parallel, we were able to discriminate three classes of defects in mitochondrial biosynthesis, namely, mitochondrial DNA depletion, mitochondrial transfer RNA (tRNA) mutations, and mutations in the mitochondrial ATP6 gene.
  
  
  CONCLUSIONS
  The relatively simple electrophoretic assay used here is a straightforward approach to differentiate between various types of genetic alterations affecting the biosynthesis of oxidative phosphorylation complexes and will be useful to guide molecular genetic diagnostics in the field of mitochondrial neuromuscular disorders.
authors.affiliation: Unit of Molecular Medicine, Bambino Gesù Hospital and Research Institute, Rome, Italy.
authors.forename: Rosalba
authors.initials: R
authors.lastname: Carrozzo
authors.forename: Ilka
authors.initials: I
authors.lastname: Wittig
authors.forename: Filippo M
authors.initials: FM
authors.lastname: Santorelli
authors.forename: Enrico
authors.initials: E
authors.lastname: Bertini
authors.forename: Sabine
authors.initials: S
authors.lastname: Hofmann
authors.forename: Ulrich
authors.initials: U
authors.lastname: Brandt
authors.forename: Hermann
authors.initials: H
authors.lastname: Schägger
mesh_terms: D001665:Binding Sites; D015415:Biomarkers; D004272:DNA, Mitochondrial / Q000378:metabolism; D042967:Electron Transport Complex I / Q000378:metabolism; D014450:Electron Transport Complex III / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D008928:Mitochondria / Q000201:enzymology* / Q000473:pathology; D028361:Mitochondrial Diseases / Q000201:enzymology* / Q000473:pathology*; D025261:Mitochondrial Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009447:Neuroblastoma; D011487:Protein Conformation; D013367:Submitochondrial Particles / Q000201:enzymology; D014407:Tumor Cells, Cultured
chemical_list: D015415:Biomarkers; D004272:DNA, Mitochondrial; C501005:MT-ATP6 protein, human; D025261:Mitochondrial Proton-Translocating ATPases; D042967:Electron Transport Complex I; D014450:Electron Transport Complex III