Mitchell 1969 Eur J Biochem
Mitchell P, Moyle J (1969) Estimation of membrane potential and pH difference across the cristae membrane of rat liver mitochondria. Eur J Biochem 7: 471-484. |
Mitchell P, Moyle J (1969) Eur J Biochem
Abstract: The cristae membrane or M phase of resting or respiring rat liver mitochondria becomes relatively permeable to K+ ions in presence of valinomycin. The equilibrium distribution of K+ ions across the membrane can therefore be used to estimate the membrane potential ΞΞ¨ provided that precautions are taken to minimise swelling of the valinomycin-treated mitochondria. The pH difference ΞpH across the M phase of anaerobic mitochondria has been estimated from the buffering powers of the inner and outer phases and from the change of pH observed on lysing the mitochondria with Triton X-100. When the anaerobic mitochondria (State 5), in presence of Ξ²-hydroxybutyrate, are brought to a state of steady respiration either in absence (State 4) or in presence (State 3) of phosphate acceptor, the changes of pH and of pK of the medium recorded with H+ ion-sensitive and K+ ion-sensitive electrodes can be used to estimate the changes of ΞΞ¨ and ΞpH across the M phase. The absolute values of ΞΞ¨ and ΞpH have been estimated from the values determined in State 5 and the change of these values in the transition from State 5 to States 4 and 3.
Continued in Free Text β’ Keywords: membrane potential, pH difference, cristae membrane, valinomycin, K+ ions
Labels: MiParea: Respiration
Organism: Rat
Tissue;cell: Liver
Preparation: Isolated Mitochondria"Isolated Mitochondria" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property.
Regulation: Coupling efficiency; uncoupling"Coupling efficiency; uncoupling" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property., Ion&substrate transport"Ion&substrate transport" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property., pH
Made history
Abstract Continued
The total protonmotive force Ξp =ΞΞ¨β 59 ΞpH across the M phase of the mitochondria oxidising Ξ²-hydroxybutyrate in State 4 at 25Β° in a 250 mM sucrose medium near pH 7 is estimated to be about 230 mV, of which the major component is ΞΞ¨ when the effect of translocation of K+ ions across the M phase is minimised. Under conditions permitting accumulation of a relatively large quantity of cation (State 6), Ξp is not significantly different from that in State 4, but the major component is βZΞpH.
The effects of changing Ξp in mitochondrial suspensions in State 4 with uncoupling agent and with pulses of acid, alkali, calcium salt and ADP have been found to be in accord with the chemiosmotic hypothesis. In particular Ξp in State 3 (ADP and Pi present) is estimated to be about 30 mV less than in State 4.