叶绿体H(+)-ATP酶二维晶体的电子冷冻显微镜观察

Electron cryomicroscopy of two-dimensional crystals of the H(+)-ATPase from chloroplasts.

作者信息

Böttcher B, Gräber P, Boekema E J, Lücken U

机构信息

Fritz-Haber Institut, Berlin, Germany.

出版信息

FEBS Lett. 1995 Oct 16;373(3):262-4. doi: 10.1016/0014-5793(95)01058-m.

DOI:10.1016/0014-5793(95)01058-m

PMID:7589479

Abstract

The H(+)-ATPase from spinach chloroplasts was isolated and purified. Two-dimensional crystals were obtained from the protein/lipid/detergent micelles by treatment with phospholipase and simultaneous removal of detergent and fatty acids by Biobeads. The resulting two-dimensionally ordered arrays were investigated by electron cryomicroscopy. The ordered arrays showed top view projections of CF0F1. The images were analysed by correlation averaging. In this view CF0F1 has dimensions of 11.4 x 9 nm. The average view shows a strongly asymmetric molecule, in contrast to the rather hexagonal features of CF1, previously analyzed from two-dimensional arrays. It is concluded that this is due either to an asymmetric structure and positioning of CF0 relative to CF1 or to a rearrangement of CF1 subunits induced by binding of CF0 to CF1.

摘要

从菠菜叶绿体中分离并纯化出H(+)-ATP酶。通过用磷脂酶处理以及利用Bio-beads同时去除去污剂和脂肪酸，从蛋白质/脂质/去污剂微团中获得二维晶体。通过电子低温显微镜对所得的二维有序阵列进行研究。有序阵列显示出CF0F1的顶视图投影。通过相关平均法对图像进行分析。在此视图中，CF0F1的尺寸为11.4×9纳米。与先前从二维阵列分析的CF1相当六边形的特征相比，平均视图显示出一个高度不对称的分子。得出的结论是，这要么是由于CF0相对于CF1的不对称结构和定位，要么是由于CF0与CF1结合诱导CF1亚基的重排。