Proton/electron stoichiometry of mitochondrial bc1 complex. Influence of pH and transmembrane delta pH

The effect of pH and transmembrane delta pH on the efficiency of the proton pump of the mitochondrial bc1 complex both in situ and in the reconstituted state was studied. In both cases the H+/e- ratio for vectorial proton translocation by the bc1 complex respiring at the steady state, under conditions in which the transmembrane pH difference (delta pH) represents the only component of the proton motive force (delta p), was significantly lower than that measured under level flow conditions. The latter amounts, at neutral pH, to 1 (2 including the scalar H+ release). In the reconstituted system steady-state delta pH was modulated by changing the intravesicular buffer as well as the intra/extra-liposomal pH. Under these conditions the H+/e- ratio varied inversely with the delta pH. The data presented show that delta pH exerts a critical control on the proton pump of the bc1 complex. Increasing the external pH above neutrality caused a decrease of the level flow H+/e- ratio. This effect is explained in terms of proton/electron linkage in b cytochromes.