Article ID Journal Published Year Pages File Type
1942674 Biochimica et Biophysica Acta (BBA) - Bioenergetics 2011 11 Pages PDF
Abstract

The metabolic control analysis was applied to digitonin-permeabilized HepG2 cell line to assess the flux control exerted by cytochrome c oxidase on the mitochondrial respiration. Experimental conditions eliciting different energy/respiratory states in mitochondria were settled. The results obtained show that the mitochondrial electrochemical potential accompanies a depressing effect on the control coefficient exhibited by the cytochrome c oxidase. Both the components of the protonmotive force, i.e. the voltage (ΔΨm) and the proton (ΔpHm) gradient, displayed a similar effect. Quantitative estimation of the ΔΨm unveiled that the voltage-dependent effect on the control coefficient of cytochrome c oxidase takes place sharply in a narrow range of membrane potential from 170–180 to 200–210 mV consistent with the physiologic transition from state 3 to state 4 of respiration. Extension of the metabolic flux control analysis to the NADH dehydrogenase and bc1 complexes of the mitochondrial respiratory chain resulted in a similar effect. A mechanistic model is put forward whereby the respiratory chain complexes are proposed to exist in a voltage-mediated threshold-controlled dynamic equilibrium between supercomplexed and isolated states.

Research highlights► MCA of mitochondrial respiration was applied to permeabilized cells. ► The mitochondrial protonmotive force depresses the flux control coefficient of COX. ► A narrow threshold range of the ΔΨmt accounts for its effect on COX. ► The respiratory flux controls of CI and CIII but not of CII respond as CIV to ΔΨmt.

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