title: Modified structure and kinetics of cytochrome-c oxidase in fibroblasts from patients with Leigh syndrome.
abstract: In this study we compared the properties of cytochrome-c oxidase (COX) in cultured fibroblasts from two patients with Leigh Syndrome with COX from control fibroblasts. The fibroblasts from patients showed decreased growth rates and elevated lactate production. COX activity of patients fibroblasts was about 25% of control. Kinetic studies with isolated mitochondria showed a higher Km for cytochrome c and a markedly reduced molecular turnover of COX from patients, indicating a different structure of the enzyme. A biphasic change of COX activity was obtained by titration of dodecylmaltoside solubilized mitochondria from control fibroblasts with increasing concentrations of anions. With patient mitochondria we found only the inhibiting phase of COX activity and, in contrast to control mitochondria, irreversible inhibition of COX activity by guanidinium chloride. ELISA titrations with monoclonal antibodies to subunit II, IV, Vab, Vlac and VIIab indicated a normal amount of mitochondrial coded subunit II, but a reduced amount of nuclear coded subunits. The data indicate incompletely assembled nuclear coded subunits of COX from patient fibroblasts.
authors.affiliation: Fachbereich Chemie, Biochemie, Philipps-Universität, Marburg, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Zimmermann
authors.forename: B
authors.initials: B
authors.lastname: Kadenbach
mesh_terms: D015153:Blotting, Western; D003902:Detergents; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D007888:Leigh Disease / Q000201:enzymology*; D008928:Mitochondria / Q000201:enzymology; D009994:Osmolar Concentration
chemical_list: D003902:Detergents; D003576:Electron Transport Complex IV