线粒体F0F1 H⁺-ATP合酶。参与H⁺转运的F0组分的特性。

Mitochondrial F0F1 H+-ATP synthase. Characterization of F0 components involved in H+ translocation.

作者信息

Guerrieri F, Capozza G, Houstĕk J, Zanotti F, Colaianni G, Jirillo E, Papa S

机构信息

Institute of Medical Biochemistry and Chemistry, Centre for the Study of Mitochondria and Energy Metabolism, CNR, Bari, Italy.

出版信息

FEBS Lett. 1989 Jun 19;250(1):60-6. doi: 10.1016/0014-5793(89)80685-4.

DOI:10.1016/0014-5793(89)80685-4

PMID:2544459

Abstract

The membrane F0 sector of mitochondrial ATP synthase complex was rapidly isolated by direct extraction with CHAPS from F1-depleted submitochondrial particles. The preparation thus obtained is stable and can be reconstituted in artificial phospholipid membranes to result in oligomycin-sensitive proton conduction, or recombined with purified F1 to give the oligomycin-sensitive F0F1-ATPase complex. The F0 preparation and constituent polypeptides were characterized by SDS-polyacrylamide gel electrophoresis and immunoblot analysis. The functional role of F0 polypeptides was examined by means of trypsin digestion and reconstitution studies. It is shown that, in addition to the 8 kDa DCCD-binding protein, the nuclear encoded protein [(1987) J. Mol. Biol. 197, 89-100], characterized as an intrinsic component of F0 (F0I, PVP protein [(1988) FEBS Lett. 237,9-14]) [corrected] is involved in H+ translocation and the sensitivity of this process to the F0 inhibitors, DCCD and oligomycin.

摘要

线粒体ATP合酶复合体的膜F0部分通过用CHAPS从去除F1的亚线粒体颗粒中直接提取而快速分离出来。由此获得的制剂是稳定的，并且可以在人工磷脂膜中重建以产生对寡霉素敏感的质子传导，或者与纯化的F1重组以得到对寡霉素敏感的F0F1 - ATP酶复合体。通过SDS - 聚丙烯酰胺凝胶电泳和免疫印迹分析对F0制剂和组成多肽进行了表征。通过胰蛋白酶消化和重建研究检查了F0多肽的功能作用。结果表明，除了8 kDa的二环己基碳二亚胺（DCCD）结合蛋白外，被表征为F0的固有成分（F0I，PVP蛋白[(1988)欧洲生物化学学会联合会快报237,9 - 14]）的核编码蛋白[（1987）《分子生物学杂志》197, 89 - 100]参与H +转运以及该过程对F0抑制剂DCCD和寡霉素的敏感性。