Brandt N R, Huber R E
Can J Biochem. 1979 Feb;57(2):145-54. doi: 10.1139/o79-018.
Differential and sucrose gradient centrifugation of honey bee thoraces, disrupted by gentle methods and using mannitol-triethanolamine-EDTA buffer at pH 6.5, showed that in the honey bee thorax 92-94.8% of the trehalase was mitochondrial. Since only 92-95% of the cytochrome c oxidase, a known mitochondrial enzyme, was found in the mitochondrial fraction by these methods, it was concluded that honey bee trehalase is totally mitochondrial. Significant amounts of 'microsomal' or 'soluble' trehalase were formed only by harsh methods of thorax disruption and similar 'microsomal' or 'soluble' trehalases were also formed by harsh treatment of purified whole mitochondria. They thus seem to be artifacts of the isolation procedure. Studies (using marker enzymes) with purified intact mitochondria which were dispersed by various chemical, enzymatic, and physical methods showed that the trehalase in the mitochondria was membrane bound and that it was bound to either the outside of the inner membrane or to one of the sides of the outer membrane.
用温和方法破坏蜜蜂胸部，并在pH 6.5的甘露醇 - 三乙醇胺 - 乙二胺四乙酸缓冲液中进行差速离心和蔗糖梯度离心，结果表明，在蜜蜂胸部，92 - 94.8%的海藻糖酶存在于线粒体中。由于通过这些方法在线粒体部分中仅发现了92 - 95%的细胞色素c氧化酶（一种已知的线粒体酶），因此得出结论，蜜蜂海藻糖酶完全存在于线粒体中。只有通过粗暴的胸部破坏方法才会形成大量的“微粒体”或“可溶性”海藻糖酶，对纯化的完整线粒体进行粗暴处理也会形成类似的“微粒体”或“可溶性”海藻糖酶。因此，它们似乎是分离过程中的人为产物。使用标记酶对通过各种化学、酶促和物理方法分散的纯化完整线粒体进行的研究表明，线粒体中的海藻糖酶与膜结合，并且它要么结合在内膜的外侧，要么结合在外膜的一侧。
