Moran O, Sciancalepore M, Sandri G, Panfili E, Bassi R, Ballarin C, Sorgato M C
Laboratorio di Neurofisiologia, Scuola Internazionale Superiore di Studi Avanzati, Trieste, Italy.
Eur Biophys J. 1992;20(6):311-9. doi: 10.1007/BF00196590.
The ionic permeability of the outer mitochondrial membrane (OMM) was studied with the patch clamp technique. Electrical recording of intact mitochondria (hence of the outer membrane (OM], derived from mouse liver, showed the presence of currents corresponding to low conductances (less than 50 pS), as well as of four distinct conductances of 99 pS, 152 pS, 220 pS and 307 pS (in 150 mM KCl). The latter were voltage gated, being open preferentially at positive (pipette) potentials. Very similar currents were found by patch clamping liposomes containing the isolated OM derived from rat brain mitochondria. Here a conductance of approximately 530 pS, resembling in its electrical characteristics a conductance already attributed to mitochondrial contact sites (Moran et al. 1990), was also detected. Immunoblot assays of mitochondria and of the isolated OM with antibodies against the outer membrane voltage-dependent anion channel (VDAC) (Colombini 1979), showed the presence of the anion channel in each case. However, the typical electrical behaviour displayed by such a channel in planar bilayers could not be detected under our experimental conditions. From this study, the permeability of the OMM appears different from what has been reported hitherto, yet is more in line with that multifarious and dynamic structure which apparently should belong to it, at least within the framework of mitochondrial biogenesis (Pfanner and Neupert 1990).
采用膜片钳技术研究了线粒体外膜(OMM)的离子通透性。对源自小鼠肝脏的完整线粒体(因而也是外膜(OM))进行电记录,结果显示存在对应于低电导(小于50 pS)的电流,以及99 pS、152 pS、220 pS和307 pS这四种不同的电导(在150 mM KCl中)。后者是电压门控的,在正(移液管)电位下优先开放。通过膜片钳技术对含有源自大鼠脑线粒体的分离外膜的脂质体进行检测,也发现了非常相似的电流。在这里,还检测到一种电导约为530 pS的电流,其电学特性类似于已归因于线粒体接触位点的一种电导(莫兰等人,1990年)。用针对外膜电压依赖性阴离子通道(VDAC)的抗体(科隆比尼,1979年)对线粒体和分离的外膜进行免疫印迹分析,结果表明每种情况下都存在阴离子通道。然而,在我们的实验条件下,未能检测到这种通道在平面双层膜中表现出的典型电学行为。从这项研究来看,线粒体外膜的通透性似乎与迄今报道的情况不同,但更符合其显然应具有的那种多样且动态的结构,至少在线粒体生物发生的框架内是如此(普法纳和诺伊佩特,1990年)。