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粟酒裂殖酵母质膜ATP酶的制备及活性测定
Preparation of Plasma Membrane ATP ases of Schizosaccharomyces pombe and Assay Their Activities
【摘要】 用小玻璃珠破碎处于对数生长中期的粟酒裂殖酵母(Schizosaccharomycespombe)细胞,将所得到的匀浆液经差速离心,酸处理,不连续蔗糖密度梯度离心,得到质膜ATP酶,并对其进行鉴定及活性测定.发现所制备的质膜ATP酶对NaN3不敏感,而对Na3VO4具有高度的敏感性,半抑制浓度为12μmol/L,表明所制备的质膜ATP酶较纯,不混杂线粒体ATP酶.该酶的最适pH为6.3,对Mg2+有很强的依赖性,对ATP有较强的亲和性,Km值为1.3mmol/L.
【Abstract】 The cells of Schizosaccharomyces pombe which were in mid-exponestial growth phase were homogenized with glass beads. The plasma membrane ATPases were obtained by differential centrifugation, followed by acid treatment and isopycnic centrifugation in a discontinuous sucrose gradient, then their activities were assayed. It was defected that the preparation of plasma membrane ATPases was insentive to NaN 3,but highly sensitive to Na 3VO 4 with an IC 50 ≈12μmol/L. It wasverified that the preparation of plasma membrane ATPases was purified and didn’t contain mitochondrial ATPases.The pH optimum of the plasma membrane ATPases was 6.3. The ATPases highly depended on Mg 2+ and showed a high affinity for ATP, Km=1.3mmol/L.
【Key words】 Schizosaccharomyces pombe; plasma membrane ATPase; preparation; assay activity;
- 【文献出处】 南京师大学报(自然科学版) ,JOURNAL OF NANJING NORMAL UNIVERSITY(NATURAL SCIENCE EDITION) , 编辑部邮箱 ,1999年02期
- 【分类号】TS261.11
- 【被引频次】2
- 【下载频次】140