SYNTHESIS AND CHARACTERIZATION OF THIOBUTYLTRIPHENYLPHOSPHONIUM BROMIDE, A NOVEL THIOL REAGENT TARGETED TO THE MITOCHONDRIAL MATRIX
SYNTHESIS AND CHARACTERIZATION OF THIOBUTYLTRIPHENYLPHOSPHONIUM BROMIDE, A NOVEL THIOL REAGENT TARGETED TO THE MITOCHONDRIAL MATRIX
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DOI：
10.1006/abbi.1995.1436
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发表时间：
1995-09-10
影响因子：
3.9
通讯作者：
MURPHY, MP
中科院分区：
文献类型：
--
作者：
BURNS, RJ;SMITH, RAJ;MURPHY, MP
由于呼吸链形成的超氧化物，线粒体持续暴露于氧化应激，其在病理情况下例如缺血再灌注和神经变性增加。在氧化应激过程中，线粒体低分子量和蛋白质巯基发生了许多变化。特别是，线粒体谷胱甘肽池被氧化并与蛋白质硫醇形成混合二硫化物。为了研究线粒体谷胱甘肽和其他线粒体巯基的氧化还原状态和共轭的变化，我们设计并表征了一种特异性靶向线粒体基质的巯基探针。该分子，硫代丁基三苯基溴化鳞，含有与亲脂性三苯基鳞阳离子连接的硫醇基团，这导致其在带负电荷的线粒体基质中积累。使用[C-14] thiobutylphenylphosphonium bromide，我们证实，它是选择性地积累分离的线粒体。在线粒体基质中，巯基与内源性巯基平衡，并在氧化应激期间与蛋白质和非蛋白质巯基形成二硫键。因此，这种新的巯基探针可用于标记蛋白质巯基，并研究在氧化应激过程中线粒体巯基的共轭和氧化还原状态的变化。(C)出版社：Academic Press
Mitochondria are continually exposed to oxidative stress due to superoxide formation by the respiratory chain which increases in pathological situations such as ischemia reperfusion and neurodegeneration. During oxidative stress there are a number of changes in mitochondrial low-molecular-weight and protein thiols. In particular, the mitochondrial glutathione pool becomes oxidized and forms mixed disulfides with protein thiols. To investigate changes in the redox state and conjugation of mitochondrial glutathione, and other mitochondrial thiols, we designed and characterized a thiol probe specifically targeted to the mitochondrial matrix. This molecule, thiobutyltriphenyl-phosphonium bromide, contains a thiol group linked to a lipophilic triphenylphosphonium cation which causes it to accumulate in the negatively charged mitochondrial matrix. Using [C-14] thiobutyltriphenylphosphonium bromide we confirmed that it was selectively accumulated by isolated mitochondria. In the mitochondrial matrix the thiol group equilibrated with endogenous thiols and during oxidative stress became disulfide-bonded to protein and nonprotein thiols. Therefore, this novel thiol probe can be used to label protein thiol groups and to investigate changes in conjugation and redox state of mitochondrial thiols during oxdative stress. (C) 1995 Academic Press, Inc.