Serviddio Gaetano, Sastre Juan
Department of Medical and Occupational Medicine, Institute of Internal Medicine, University of Foggia, Foggia, Italy.
Methods Mol Biol. 2010;594:107-21. doi: 10.1007/978-1-60761-411-1_7.
The major component of mitochondrial electrochemical potential gradient of protons is the mitochondrial membrane potential (DeltaPsi), and hence it is a suitable parameter for assessment of mitochondrial function. Dissipation of the mitochondrial membrane potential causes uncoupling of the electron transport through the respiratory chain and the phosphorylation reaction for ATP synthesis (proton leak). Proton leak functions as a regulator of mitochondrial reactive oxygen species (ROS) production and its modulation by uncoupling proteins, which may be involved in pathophysiology. In this report, we describe the assays for mitochondrial membrane potential and proton leak, which require a TPP(+) electrode and a Clark electrode. The determination of mitochondrial peroxide production with homovanillic acid is also included.
质子线粒体电化学势梯度的主要成分是线粒体膜电位（ΔΨ），因此它是评估线粒体功能的一个合适参数。线粒体膜电位的耗散会导致通过呼吸链的电子传递与ATP合成的磷酸化反应解偶联（质子泄漏）。质子泄漏作为线粒体活性氧（ROS）产生的调节剂，并通过解偶联蛋白对其进行调节，这可能参与病理生理学过程。在本报告中，我们描述了线粒体膜电位和质子泄漏的检测方法，这些方法需要一个四苯硼钠（TPP(+)）电极和一个克拉克电极。同时还包括用高香草酸测定线粒体过氧化物的产生。
