Gorneva G A, Skopinskaia S N, Demin V V, Riabova I D
Biokhimiia. 1976 Jul;41(6):1033-7.
Membrane fractions were isolated from Streptococcus faecalis cells of a glycolyzing microorganism, devoid of the respiratory chain, using the methods of osmotic shock of the protoplasts, ultrasonic treatment of the cells and ultrasonic treatment of the protoplasts. All fractions possessed the ATPase activity, the highest activity being observed in the fraction isolated by ultrasonication of the protoplasts. All preparations were estimated with respect to the presence of vesicles, formed by the "inside-out" and "inside-in" membranes, using ATPase as a marker of the membrane orientation. In the membrane fractions obtained by ultrasonication of the protoplasts, the "inside-out" vesicles were prevalent. ATP-dependent energization of the membranes, sensitive to the action of dicyclohexylcarbodiimide and tetrachlorotrifluoromethyl benzimidazole, was demonstrated by measuring the transport of the lipophylic anion of phenyldicarbaundecaborane and aniline naphthalene sulfonate fluorescence.
使用原生质体渗透休克法、细胞超声处理法和原生质体超声处理法,从缺乏呼吸链的糖酵解微生物粪肠球菌细胞中分离出膜组分。所有组分都具有ATP酶活性,其中通过原生质体超声处理分离得到的组分中观察到的活性最高。使用ATP酶作为膜取向的标志物,对所有制剂中由“外翻”和“内翻”膜形成的囊泡的存在情况进行了评估。在通过原生质体超声处理获得的膜组分中,“外翻”囊泡占主导。通过测量苯基二碳十一硼烷的亲脂性阴离子的转运和苯胺萘磺酸盐荧光,证明了膜的ATP依赖性能量化对二环己基碳二亚胺和四氯三氟甲基苯并咪唑的作用敏感。