Purification, Reconstitution and H+ Translocation Activity of Porcine Heart Mitochondrial Fo
- VernacularTitle:猪心线粒体Fo的纯化、重建及其质子转运功能
- Author:
Hui YANG
;
Ling ZHANG
;
Xujia ZHANG
;
Youguo HUANG
- Publication Type:Journal Article
- Keywords:
submitochondria;
Fo;
reconstitution;
H+ translocati on
- From:
Progress in Biochemistry and Biophysics
2001;28(2):227-231
- CountryChina
- Language:Chinese
-
Abstract:
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.
