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 .

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.