Klingenberg M, Rottenberg H
Eur J Biochem. 1977 Feb 15;73(1):125-30. doi: 10.1111/j.1432-1033.1977.tb11298.x.
The relation between the intramitochondrial and extramitochondrial ratio ATP/ADP, the transmembrane potential and pH gradient is investigated in the present communication. For this purpose mitochondria are equilibrated with added [14C]ATP in the presence of substrate and oligomycin for eliminating phosphate transfer by ATPase. The membrane potential was measured by the distribution of 86Rb+ in the presence of valinomycin, the deltapH by the distribution of [14C]acetate. In the energized state by varying deltapsi between 60 and 160 mV, the internal (ATP/ADP)i is decreased 30-fold, the external (ATP/ADP)e remains largely constant. As a result, the deltalog (ATP/ADP)e/(ATP/ADP)i = deltalogphi is increased linerly with deltapsi according to the following relation: deltalogphi = 0.85 deltapsi - 0.35. The deltapH was changed between 0.1 and 0.8 by increasing the Pi concentration causing only a minor decrease of deltalogphi would be expected if the ATP-ADP exchange has a significant electroneutral portion. Also in the uncoupled and respiration-inhibited state the same function between deltalogphi and deltapsi is found as in the energized states. It is concluded that under these conditions the ATP-ADP exchange is largely electrical.
本文研究了线粒体内外ATP/ADP比值、跨膜电位和pH梯度之间的关系。为此,在底物和寡霉素存在的情况下,用添加的[14C]ATP使线粒体达到平衡,以消除ATP酶介导的磷酸转移。通过在缬氨霉素存在下86Rb+的分布来测量膜电位,通过[14C]乙酸盐的分布来测量ΔpH。在能量化状态下,通过将Δψ在60至160 mV之间变化,内部(ATP/ADP)i降低30倍,外部(ATP/ADP)e基本保持恒定。结果,Δlog(ATP/ADP)e/(ATP/ADP)i = Δlogφ根据以下关系随Δψ线性增加:Δlogφ = 0.85Δψ - 0.35。通过增加Pi浓度使ΔpH在0.1至0.8之间变化,如果ATP-ADP交换具有显著的电中性部分,则只会导致Δlogφ略有下降。在解偶联和呼吸抑制状态下,也发现Δlogφ和Δψ之间的函数关系与能量化状态相同。得出的结论是,在这些条件下,ATP-ADP交换主要是电性质的。