Burns R J, Murphy M P
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
Arch Biochem Biophys. 1997 Mar 1;339(1):33-9. doi: 10.1006/abbi.1996.9861.
Alterations to the redox state of mitochondrial thiols is a particularly important response to oxidative stress, because mitochondria are a major source of reactive oxygen species within cells. To investigate these changes we designed and synthesized a novel probe for mitochondrial thiols, thiobutyltriphenylphosphonium bromide [TBTP; R. J. Burns, R. A. J. Smith, and M. P. Murphy (1995) Arch. Biochem. Biophys. 322, 60-68]. This lipophilic cation was accumulated several hundred-fold into the negatively charged matrix of isolated mitochondria, where it equilibrated with endogenous thiols and during oxidative stress formed disulfide bonds to exposed protein thiols. In this paper we show that TBTP can also selectively react with mitochondrial protein thiols in living cells. Since TBTP localizes specifically to mitochondria and forms disulfide bonds selectively with mitochondrial proteins during oxidative stress, we conclude that TBTP has utility for investigating changes in mitochondrial thiols, independently of alterations occurring in the cytoplasm.
线粒体硫醇氧化还原状态的改变是对氧化应激的一种特别重要的反应，因为线粒体是细胞内活性氧的主要来源。为了研究这些变化，我们设计并合成了一种用于线粒体硫醇的新型探针，硫代丁基三苯基溴化膦[TBTP；R. J. 伯恩斯、R. A. J. 史密斯和M. P. 墨菲（1995年）《生物化学与生物物理学报》322卷，60 - 68页]。这种亲脂性阳离子被数百倍地积累到分离线粒体带负电荷的基质中，在那里它与内源性硫醇平衡，并在氧化应激期间与暴露的蛋白质硫醇形成二硫键。在本文中，我们表明TBTP也能在活细胞中与线粒体蛋白质硫醇选择性反应。由于TBTP特异性定位于线粒体，并在氧化应激期间与线粒体蛋白质选择性形成二硫键，我们得出结论，TBTP可用于研究线粒体硫醇的变化，而与细胞质中发生的变化无关。
