title: Flux control of cytochrome c oxidase in human skeletal muscle.
abstract: In the present work, by titrating cytochrome c oxidase (COX) with the specific inhibitor KCN, the flux control coefficient and the metabolic reserve capacity of COX have been determined in human saponin-permeabilized muscle fibers. In the presence of the substrates glutamate and malate, a 2.3 +/- 0.2-fold excess capacity of COX was observed in ADP-stimulated human skeletal muscle fibers. This value was found to be dependent on the mitochondrial substrate supply. In the combined presence of glutamate, malate, and succinate, which supported an approximately 1.4-fold higher rate of respiration, only a 1.4 +/- 0.2-fold excess capacity of COX was determined. In agreement with these findings, the flux control of COX increased, in the presence of the three substrates, from 0.27 +/- 0.03 to 0.36 +/- 0.08. These results indicate a tight in vivo control of respiration by COX in human skeletal muscle. This tight control may have significant implications for mitochondrial myopathies. In support of this conclusion, the analysis of skeletal muscle fibers from two patients with chronic progressive external ophthalmoplegia, which carried deletions in 11 and 49% of their mitochondrial DNA, revealed a substantially lowered reserve capacity and increased flux control coefficient of COX, indicating severe rate limitations of oxidative phosphorylation by this enzyme.
authors.affiliation: Department of Epileptology, University Bonn Medical Center, Sigmund-Freud-Strasse 25, D-53105 Bonn, Germany. kunz@mailer.meb.uni-bonn.de
authors.forename: W S
authors.initials: WS
authors.lastname: Kunz
authors.forename: A
authors.initials: A
authors.lastname: Kudin
authors.forename: S
authors.initials: S
authors.lastname: Vielhaber
authors.forename: C E
authors.initials: CE
authors.lastname: Elger
authors.forename: G
authors.initials: G
authors.lastname: Attardi
authors.forename: G
authors.initials: G
authors.lastname: Villani
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000328:Adult; D015139:Blotting, Southern; D002463:Cell Membrane Permeability; D004272:DNA, Mitochondrial / Q000235:genetics; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008875:Middle Aged; D008931:Mitochondria, Muscle / Q000201:enzymology; D017240:Mitochondrial Myopathies / Q000201:enzymology; D018485:Muscle Fibers, Skeletal / Q000201:enzymology / Q000473:pathology; D018482:Muscle, Skeletal / Q000201:enzymology* / Q000473:pathology; D017246:Ophthalmoplegia, Chronic Progressive External / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D010101:Oxygen Consumption / Q000187:drug effects; D011190:Potassium Cyanide / Q000494:pharmacology*; D017384:Sequence Deletion
chemical_list: D004272:DNA, Mitochondrial; D000244:Adenosine Diphosphate; D003576:Electron Transport Complex IV; D011190:Potassium Cyanide