Abstract
The F0 membrane domain of F0F1-ATPase complex had been purified from porcine heart mitochondria. SDS-PAGE with silver staining indicated that the purity of F0 was about 85% and the sample contained no subunits of F1-ATPase. The purified F0 was reconstituted into liposomes with different phospholipid composition, and the effect of CL (cardiolipin), PA (phosphatidic acid), PI (phosphatidylinositol) and PS (phosphatidylserine) on the H+ translocation activity of F0 was investigated. The results demonstrated that CL, PA and PI could promote the proton translocation of F0 with the order of CLPA>PI, while PS inhibited it. Meanwhile ADM (adriamycin) severely impaired the proton translocation activity of F0 vesicles containing CL, which suggested that CL’s stimulation of the activity of reconstituted F0 might correlate with its non-bilayer propensity. After F0 was incorporated into the liposomes containing PE (phosphatidylethanolamine), DOPE (dioleoylphosphatidylethanolamine) as well as DEPE (dielaidoylphosphatidylethanolamine), it was found that the proton translocation activity of F0 vesicles increased with the increasing content of PE or DOPE, which has high propensity of forming non-bilayer structure, but was independent of DEPE. The dynamic quenching of the intrinsic fluorescence of tryptophan by HB (hypocrellin B) as well as fluorescent spectrum of acrylodan labeling F0 at cysteine indicated that CL could induce F0 to a suitable conformation resulting in higher proton translocation activity.
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Yang, H., Huang, Y., Zhang, X. et al. Cardiolipin is essential for higher proton translocation activity of reconstituted F0 . Sci. China Ser. C.-Life Sci. 44, 146–155 (2001). https://doi.org/10.1007/BF02879319
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DOI: https://doi.org/10.1007/BF02879319