猪心线粒体F_o的纯化、重建及其质子转运功能

比较了猪心线粒体FoF1-ATPase膜部分Fo的四种纯化方法 .结果表明，用NaBr从亚线粒体除去FoF1-ATPase的水溶性部分F1-ATPase后，再以C HAPS增溶，并经蔗糖梯度离心，可获得高纯度的Fo.SDS-聚丙烯酰胺凝胶电泳鉴定表明， 纯化的Fo含有b、OSCP（寡霉素敏感授予蛋白）、d、a、e、F6、IF1、A6L和c等9种亚 基.用去污剂稀释法将纯化的Fo在脂质体上重建后，重建Fo表现较高的被动转运质子活 性. 这为在体外深入研究Fo的活性、构象与膜脂的关系，以及Fo与F1-ATPase的组装等提 供了很好的实验模型.

Purification, Reconstitution and H~ Translocation Activity of Porcine Heart Mitochondrial F_o

Four methods were compared to purify Fo from porcine h ear t mitochondria. The best results were obtained by the following method: after re moving F1-ATPase with NaBr incubation from submitochondrial FoF1-ATPase, Fo was solubilized with CHAPS and purified by sucrose density gradient centri fugation. SDS-PAGE with silver staining showed about 85% purity of the isolated Fo and 9 different subunits including b, OSCP, d, a, e, F6, IF1, A6L and c. The purified Fo was then incorporated into asolectin liposomes, the reconst ituted Fo showed higher H+ translocation activity and after Fo was reconst ituted with F1-ATPase, the resulted FoF1-ATPase complex exhibited high A TP hydrolysis activity and high sensitivity to oligomycin. The results provide e vidence for successful purification, reconstitution of Fo with high H+ trans location activity and its relationship with phospholipids.