胡杨液泡膜H+-ATPase的部分纯化及其耐盐性研究

为了阐明液泡膜H^ -ATPase在盐胁迫下的作用和适应性机制，对悬浮培养的胡杨细胞在50mmol／L盐浓度下处理10d，结果表明液泡膜H^ -ATPase、焦磷酸酶的水解活性、质子泵活性增加。将通过差速离心和不连续蔗糖密度梯度离心富积的液泡膜微囊先由脱氧胆酸钠(DOC)和n-辛基-β-D-葡萄糖(OG)分步破膜抽提，经蔗糖密度梯度离心分离，部分纯化的酶含V型H^ -ATPase的主要亚基。

Studies on purification of tonoplast H+-ATPase from Populus euphratica and its salt tolerance

To study the function and adaptive mechanism of tonoplast H~+-ATPase under salt stress,Populus euphratica suspension cultured cells were treated with 50 mmol/L NaCl for 10 days,the ATP,PPiase hydrolytic activity and the proton transport activity increased under the salt stress.Tonoplast-enriched vesicles by differential centrifugation and discontinuous sucrose density gradient centrifugation were first treated by 0.15% DOC to remove other membrane proteins.The H~+-ATPase still on the membrane was further solubilized by 40 mmol/L OG,the partially purified enzyme through sucrose density gradients centrifugation contained polypeptides of apparent molecular mass 68,57,30,26 and 17 kD resaved by SDS-PAGE.They were similar to the subunits of H~+-ATPase from other sources.