Mitochondrial respiration at low levels of oxygen and cytochrome c.
In the intracellular microenvironment of active muscle tissue, high rates of respiration are maintained at near-limiting oxygen concentrations. The respiration of isolated heart mitochondria is a hyperbolic function of oxygen concentration and half-maximal rates were obtained at 0.4 and 0.7 microM O(2) with substrates for the respiratory chain (succinate) and cytochrome c oxidase [N,N,N,N',N'-tetramethyl-p-phenylenediamine dihydrochloride (TMPD)+ascorbate] respectively at 30 degrees C and with maximum ADP stimulation (State 3). The respiratory response of cytochrome c-depleted mitoplasts to external cytochrome c was biphasic with TMPD, but showed a monophasic hyperbolic function with succinate. Half-maximal stimulation of respiration was obtained at 0.4 microM cytochrome c, which was nearly identical to the high-affinity K(')(m) for cytochrome c of cytochrome c oxidase supplied with TMPD. The capacity of cytochrome c oxidase in the presence of TMPD was 2-fold higher than the capacity of the respiratory chain with succinate, measured at environmental normoxic levels. This apparent excess capacity, however, is significantly decreased under physiological intracellular oxygen conditions and declines steeply under hypoxic conditions. Similarly, the excess capacity of cytochrome c oxidase declines with progressive cytochrome c depletion. The flux control coefficient of cytochrome c oxidase, therefore, increases as a function of substrate limitation of oxygen and cytochrome c, which suggests a direct functional role for the apparent excess capacity of cytochrome c oxidase in hypoxia and under conditions of intracellular accumulation of cytochrome c after its release from mitochondria.
Gnaiger
Erich
E
Department of Transplant Surgery, D. Swarovski Research Laboratory, University Hospital Innsbruck, Anichstrasse 35, A-6020 Innsbruck, Austria. erich.gnaiger@uibk.ac.at
Kuznetsov
A V
AV
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003574:Cytochrome c Group / Q000378:metabolism*; D003576:Electron Transport Complex IV / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008929:Mitochondria, Heart / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption; D051381:Rats; D011917:Rats, Inbred Lew; D019802:Succinic Acid / Q000378:metabolism; D013772:Tetramethylphenylenediamine / Q000378:metabolism
D003574:Cytochrome c Group; D000255:Adenosine Triphosphate; D019802:Succinic Acid; D003576:Electron Transport Complex IV; D013772:Tetramethylphenylenediamine; D010100:Oxygen