嗜热栖热菌变构L-乳酸脱氢酶的结晶及初步晶体学数据。

Crystallization of allosteric L-lactate dehydrogenase from Thermus caldophilus and preliminary crystallographic data.

作者信息

Koide S, Iwata S, Matsuzawa H, Ohta T

机构信息

Department of Agricultural Chemistry, University of Tokyo.

出版信息

J Biochem. 1991 Jan;109(1):6-7.

PMID:2016275

Abstract

L-Lactate dehydrogenase of Thermus caldophilus GK24 was purified from Escherichia coli containing an overexpression plasmid. The enzyme was crystallized from polyethylene glycol 6000 solutions without ligands by the hanging drop vapor diffusion method. Two forms of crystals were obtained. The crystals grown at pH 6.0 were characterized by means of an X-ray diffraction experiment, while those grown at pH 6.5 and 7.0 did not give detectable diffraction spots. The crystals grown at pH 6.0 belonged to monoclinic space group P2(1), the cell dimensions being a = 54.8 A, b = 138.2A, c = 86.1 A, and beta = 93.3 degrees. These crystals diffract to beyond 2.5 A spacing and are stable on X-ray irradiation.

摘要

嗜热栖热菌GK24的L-乳酸脱氢酶是从含有过表达质粒的大肠杆菌中纯化得到的。通过悬滴气相扩散法，该酶在无配体的聚乙二醇6000溶液中结晶。获得了两种晶体形式。在pH 6.0条件下生长的晶体通过X射线衍射实验进行表征，而在pH 6.5和7.0条件下生长的晶体未给出可检测到的衍射斑点。在pH 6.0条件下生长的晶体属于单斜空间群P2(1)，晶胞参数为a = 54.8 Å，b = 138.2 Å，c = 86.1 Å，β = 93.3°。这些晶体的衍射分辨率超过2.5 Å间距，并且在X射线照射下稳定。