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大肠杆菌纤维二糖特异性磷酸转移酶系统中酶IIB的结晶

Crystallization of enzyme IIB of the cellobiose-specific phosphotransferase system of Escherichia coli.

作者信息

van Montfort R L, Pijning T, Kalk K H, Schuurman-Wolters G K, Reizer J, Saier M H, Robillard G, Dijkstra B W

机构信息

Laboratory of Biophysical Chemistry, University of Groningen, The Netherlands.

出版信息

J Mol Biol. 1994 Jun 17;239(4):588-90. doi: 10.1006/jmbi.1994.1399.

DOI:10.1006/jmbi.1994.1399
PMID:8006971
Abstract

Crystals of enzyme IIB of the cellobiose-specific phosphotransferase system have been obtained from 15% polyethylene glycol 4000 using both streak-seeding and macroseeding techniques at 4 degrees. Crystals were grown with the hanging drop method of vapour diffusion. Addition of 2-propanol and benzamidine/HCl proved essential to obtain single crystals suitable for X-ray analysis. The crystals diffract to 1.8 A resolution and have the monoclinic space group P2(1), with cell dimensions a = 53.6 A, b = 31.7 A, c = 60.0 A and beta = 101.7 degrees. From a self-rotation function it seems likely that there are two molecules in the asymmetric unit related by a non-crystallographic 2-fold axis.

摘要

已通过在4℃下使用划线接种和宏观接种技术,从15%的聚乙二醇4000中获得了纤维二糖特异性磷酸转移酶系统的酶IIB晶体。晶体采用气相扩散悬滴法生长。事实证明,添加2-丙醇和盐酸苯甲脒对于获得适合X射线分析的单晶至关重要。这些晶体的衍射分辨率为1.8埃,属于单斜空间群P2(1),晶胞参数为a = 53.6埃,b = 31.7埃,c = 60.0埃,β = 101.7°。从自旋转函数来看,不对称单元中可能有两个通过非晶体学2重轴相关的分子。

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