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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

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【作者】 林玲袁生幸定坤陆玲张如海

【Author】 Lin Ling,Yuan Sheng,Xing Dingkun,Lu Ling,Zhang Ruhai Department of Biology, Nanjing Normal University, 210097,Nanjing, PRC.

【机构】 南京师范大学生物系

【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.

【基金】 江苏省自然科学基金
  • 【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
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