Fedotov N S, Panchenko L F, Logachev A P, Bekkouzhin A G, Tarshis M A
Folia Microbiol (Praha). 1975;20(6):470-9. doi: 10.1007/BF02891706.
A new procedure for the isolation of membrane vesicles from Acholeplasma laidlawii cells is described. The membrane vesicles are completely free from contaminations of whole cells and cell debris and represent a homogeneous fraction as shown by electron microscopy, Ficoll density-gradient centrifugation, and titration on agar plate. Absence of cytoplasmic contaminations was confirmed by double-labelling of membranes with 3H-oleic acid and 14C-uridine, as well as by distribution of specific marker enzymes of membranes and cytoplasm. On the basis of light-scattering and electron microscopy, the vesicular nature of these membranes was established. The vesicles had the same orientation as intact cells (absence on membrane vesicles of ATPase and NADH dehydrogenase activities, localized in the inner surface of membrane). The respiratory activity of the membrane vesicles was low and was not stimulated by exogenous substrates, the respiratory chain of the vesicles being reduced and terminated by flavoproteins. The ability of membrane vesicles to take up carbohydrates was shown.
本文描述了一种从莱氏无胆甾原体细胞中分离膜泡的新方法。膜泡完全没有全细胞和细胞碎片的污染，并且通过电子显微镜、菲可密度梯度离心和琼脂平板滴定显示为均一的组分。通过用³H-油酸和¹⁴C-尿苷对膜进行双重标记，以及通过膜和细胞质特异性标记酶的分布，证实了不存在细胞质污染。基于光散射和电子显微镜，确定了这些膜的泡状性质。这些膜泡与完整细胞具有相同的取向（膜泡上不存在定位于膜内表面的ATP酶和NADH脱氢酶活性）。膜泡的呼吸活性较低，并且不受外源底物的刺激，膜泡的呼吸链被黄素蛋白还原并终止。研究表明膜泡具有摄取碳水化合物的能力。
