嗜热芽孢杆菌PS3的F1-ATP酶突变β亚基的结晶

Crystallization of mutant beta subunit of F1-ATPase from thermophilic Bacillus PS3.

作者信息

Saika K, Inaka K, Matsui T, Yoshida M, Miki K

机构信息

Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan.

出版信息

J Mol Biol. 1994 Oct 7;242(5):709-11. doi: 10.1006/jmbi.1994.1621.

DOI:10.1006/jmbi.1994.1621

PMID:7932728

Abstract

The mutant beta subunit of F1-ATPase from a thermophilic Bacillus strain, PS3, in which tyrosine at position 341 is replaced by leucine (beta Y341L) was expressed in Escherichia coli and crystallized by the vapor-diffusion procedure. Small needle-like crystals were obtained using ammonium sulfate as a precipitant and grown by the stepwise seeding method. The crystals obtained by this procedure diffracted X-rays to about 3 A resolution. The diffraction patterns indicated that the crystals belong to the orthorhombic system and the space group I222 or I2(1)2(1)2(1) with unit-cell dimensions of a = 232 A, b = 66 A, and c = 80 A. It is thought that the asymmetric unit comprises one beta Y341L molecule.

摘要

来自嗜热芽孢杆菌菌株PS3的F1 - ATP酶突变β亚基，其中第341位的酪氨酸被亮氨酸取代（βY341L），在大肠杆菌中表达，并通过气相扩散法结晶。使用硫酸铵作为沉淀剂获得了小针状晶体，并通过逐步接种法生长。通过该方法获得的晶体将X射线衍射到约3埃的分辨率。衍射图谱表明，这些晶体属于正交晶系，空间群为I222或I2(1)2(1)2(1)，晶胞参数为a = 232埃，b = 66埃，c = 80埃。据认为，不对称单元包含一个βY341L分子。