胡杨质膜的纯化及其H^+-ATPase活性的研究  

Study on Purification and Activity of Plasma Membrane H^+-ATPase of Populus euphratica .

作　　者：张玲[1] 刘群录 黄有国[1] 

机构地区：[1]中国科学院生物物理研究所 [2]北京中国林业大学生物学院,北京100083

出　　处：《生物化学与生物物理进展》2000年第6期637-640,共4页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金重点项目基金支持!(39830 32 0 ;39730 130 )

摘　　要：用DextranT 5 0 0 ,PEG 335 0两相分配法分离并纯化了悬浮培养的胡杨细胞质膜 .不同聚合物浓度(5 5 %、 5 7%、 5 9%、 6 1%、 6 3%、 6 5 % )和KCl浓度 (0、 5、 10、 15mmol/L)对分离效果影响的研究结果表明 ,采用聚合物浓度为 5 9%和无盐存在的两相分配体系可获得纯度较高的胡杨细胞质膜 .纯化的质膜H+ ATPase的活力提高 8倍 ,且酶定向程度较高 ,这为进一步研究胡杨细胞质膜特性及获得高纯度H+The plasma membrane from suspended cultured Populus euphratica (PE) cells had been isolated and purified by two phase partition composed of Dextran T 500 and PEG 3350. Effect of different concentrations of the polymer (5 5%,5 7%,5 9%,6 1%,6 3%,6 5%) and KCl (0,5,10,15 mmol/L) on the purification were examined. The results indicated that two phase system composed of 5 9% polymer without salt resulted in plasma membrane of PE with higher H + ATPase activity. The activity of the membrane H + ATPase has been increased 8 fold (from 4 23 to 32 63 μmol/mg·h) with better orientation and most of the membrane H + ATPases are right side out. These results may provide better basis and availability for further studies of the purified H + ATPase from PE plasma membrane.

关 键 词：质膜 两相分配法 H^+-ATPASE 纯化 细胞质膜 

分 类 号：Q73[生物学—分子生物学] Q942