生物珠：一种用于膜蛋白二维结晶的有效策略。

Bio-Beads: an efficient strategy for two-dimensional crystallization of membrane proteins.

作者信息

Rigaud J L, Mosser G, Lacapere J J, Olofsson A, Levy D, Ranck J L

机构信息

Section de recherche, Institut Curie, UMR-CNRS 168 and LRC-CEA 8, Paris, France.

出版信息

J Struct Biol. 1997 Apr;118(3):226-35. doi: 10.1006/jsbi.1997.3848.

DOI:10.1006/jsbi.1997.3848

PMID:9169232

Abstract

This work establishes the potential of Bio-Beads as a simple alternative to conventional dialysis for removing detergent and for obtaining 2D crystals of integral membrane proteins useful for structure analysis by electron crystallography. Kinetic and equilibrium aspects of removal of different detergents by adsorption onto hydrophobic Bio-Beads SM2 have been systematically investigated and extended to 2D crystallization of different prototypic membrane proteins, including: (a) Ca2+ ATPase from sarcoplasmic reticulum; (b) melibiose permease from Escherichia coli; (c) cytochrome b6f from Chlamydomonas reinhardtii. Different crystals could be produced from all protein preparations, with optical diffraction down to 20-25 A in negative stain.

摘要

这项工作证实了Bio-Beads作为一种简单替代方法的潜力，可用于去除去污剂以及获得用于电子晶体学结构分析的完整膜蛋白二维晶体。通过吸附到疏水Bio-Beads SM2上，对去除不同去污剂的动力学和平衡方面进行了系统研究，并扩展到不同原型膜蛋白的二维结晶，包括：(a) 肌浆网Ca2+ ATP酶；(b) 大肠杆菌蜜二糖通透酶；(c) 莱茵衣藻细胞色素b6f。所有蛋白质制剂都能产生不同的晶体，在负染下光学衍射低至20-25埃。