Deryabina Y I, Isakova E P, Shurubor E I, Zvyagilskaya R A
Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, 119071, Russia.
Biochemistry (Mosc). 2004 Sep;69(9):1025-33. doi: 10.1023/b:biry.0000043545.32343.50.
Mitochondria of the yeast Endomyces magnusii were examined for the presence of a Ca2+- and phosphate-induced permeability of the inner mitochondrial membrane (pore). For this purpose, coupled mitochondria were incubated under conditions known to induce the permeability transition pore in animal mitochondria, i.e., in the presence of high concentrations of Ca2+ and P(i), prooxidants (t-butylhydroperoxide), oxaloacetate, atractyloside (an inhibitor of ADP/ATP translocator), SH-reagents, by depletion of adenine nucleotide pools, and deenergization of the mitochondria. Large amplitude swelling, collapse of the membrane potential, and efflux of the accumulated Ca2+ were used as parameters for demonstrating pore induction. E. magnusii mitochondria were highly resistant to the above-mentioned substances. Deenergization of mitochondria or depletion of adenine nucleotide pools have no effect on low-amplitude swelling or the other parameters. Cyclosporin A, a specific inhibitor of the nonspecific permeability transition in animal mitochondria, did not affect the parameters measured. It is thus evident that E. magnusii mitochondria lack a functional Ca2+-dependent pore, or possess a pore differently regulated as compared to that of mammalian mitochondria.
对大孢内孢霉酵母的线粒体进行了检查,以确定线粒体内膜是否存在Ca2+和磷酸盐诱导的通透性(孔道)。为此,将偶联的线粒体在已知能诱导动物线粒体通透性转换孔道的条件下孵育,即在高浓度Ca2+和无机磷酸(Pi)、促氧化剂(叔丁基过氧化氢)、草酰乙酸、苍术苷(ADP/ATP转位酶抑制剂)、巯基试剂存在的情况下,通过消耗腺嘌呤核苷酸池以及使线粒体去能。大幅度肿胀、膜电位崩溃和积累的Ca2+外流被用作证明孔道诱导的参数。大孢内孢霉酵母线粒体对上述物质具有高度抗性。线粒体去能或腺嘌呤核苷酸池的消耗对低幅度肿胀或其他参数没有影响。环孢素A是动物线粒体非特异性通透性转换的特异性抑制剂,对所测参数没有影响。因此很明显,大孢内孢霉酵母线粒体缺乏功能性的Ca2+依赖性孔道,或者拥有与哺乳动物线粒体不同调节方式的孔道。