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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >The effect of bovine serum albumin on the membrane potential and reactive oxygen species generation in succinate-supported isolated brain mitochondria.
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The effect of bovine serum albumin on the membrane potential and reactive oxygen species generation in succinate-supported isolated brain mitochondria.

机译：牛血清白蛋白对琥珀酸支持的离体脑线粒体膜电位和活性氧生成的影响。

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摘要

Characteristics of the succinate-supported H(2)O(2) formation were compared in mitochondria prepared from guinea-pig brain either by Percoll gradient centrifugation or using digitonin. The high rate of H(2)O(2) generation measured in mitochondria prepared with digitonin (600.6+/-26.8pmol/min/mg protein) was inhibited by rotenone, consistently with a reverse flow of electrons via complex I. The rate of H(2)O(2) formation was significantly smaller in Percoll-purified mitochondria (252.6+/-17.3pmol/min/mg protein) and this was stimulated by rotenone. Since bovine serum albumin (BSA) is usually present in the isolation medium used in the digitonin method, systematic study was performed addressing the effect of BSA on H(2)O(2) formation. Mitochondria prepared by the digitonin method (BSA present in the isolation medium) were highly polarized (185+/-3.2mV) and addition of BSA (0.025%) to the assay medium increased H(2)O(2) generation by only 50%. In Percoll-purified mitochondria DeltaPsim was more depolarized (171+/-2mV) and BSA caused hyperpolarization by 10.7+/-1.9mV. H(2)O(2) formation, which was largely independent of DeltaPsim, was stimulated by 400%, became highly dependent on DeltaPsim and could be inhibited by rotenone in the presence of BSA. This shows that in Percoll-purified mitochondria ROS formation via reverse electron flow is preferred only when BSA is present in the assay medium. It is demonstrated that (i) the presence or absence of BSA could determine the mechanism by which ROS is generated in succinate-supported mitochondria and (ii) depolarization by about 10mV eliminates reverse electron flow and the remaining ROS formation, which is smaller but still significant, is no longer dependent on DeltaPsim.
机译：琥珀酸支持的H（2）O（2）形成的特征在通过Percoll梯度离心法或使用洋地黄皂苷从豚鼠脑制备的线粒体中进行了比较。鱼藤酮抑制了高剂量H（2）O（2）生成的线粒体中的洋地黄酮（600.6 +/- 26.8pmol / min / mg蛋白质）的生成，与通过复合体I反向流动的电子一致。 Percoll纯化的线粒体中的H（2）O（2）形成的比例显着较小（252.6 +/- 17.3pmol / min / mg蛋白质），并且受到鱼藤酮的刺激。由于牛血清白蛋白（BSA）通常存在于洋地黄皂苷方法中使用的分离介质中，因此进行了系统的研究，以解决BSA对H（2）O（2）形成的影响。通过指骨蛋白法制备的线粒体（分离介质中存在BSA）被高度极化（185 +/- 3.2mV），向测定介质中添加BSA（0.025％）仅使H（2）O（2）生成增加了50 ％。在Percoll纯化的线粒体中，DeltaPsim的去极化程度更高（171 +/- 2mV），而BSA引起的超极化程度为10.7 +/- 1.9mV。 H（2）O（2）的形成，很大程度上独立于DeltaPsim，被400％刺激，变得高度依赖DeltaPsim，并可能在存在BSA的情况下被鱼藤酮抑制。这表明在Percoll纯化的线粒体中，仅当测定介质中存在BSA时，才优选通过反向电子流形成ROS。已证明（i）是否存在BSA可以决定在琥珀酸酯负载的线粒体中产生ROS的机制，以及（ii）约10mV的去极化消除了反向电子流，并消除了残留的ROS形成，后者较小但仍然重要，不再依赖于DeltaPsim。

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