CAJ | 학술논문

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.
비교료저심선립체FoF1-ATPase막부분Fo적사충순화방법 .결과표명，용NaBr종아선립체제거FoF1-ATPase적수용성부분F1-ATPase후，재이C HAPS증용，병경자당제도리심，가획득고순도적Fo.SDS-취병희선알응효전영감정표명， 순화적Fo함유b、OSCP（과매소민감수여단백）、d、a、e、F6、IF1、A6L화c등9충아 기.용거오제희석법장순화적Fo재지질체상중건후，중건Fo표현교고적피동전운질자활 성. 저위재체외심입연구Fo적활성、구상여막지적관계，이급Fo여F1-ATPase적조장등제 공료흔호적실험모형.
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.