大肠杆菌K-12铜胺氧化酶的结晶及初步X射线分析

Crystallization and preliminary X-ray analysis of copper amine oxidase from Escherichia coli K-12.

作者信息

Roh J H, Suzuki H, Kumagai H, Yamashita M, Azakami H, Murooka Y, Mikami B

机构信息

Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.

出版信息

J Mol Biol. 1994 May 13;238(4):635-7. doi: 10.1006/jmbi.1994.1321.

DOI:10.1006/jmbi.1994.1321

PMID:8176752

Abstract

Copper-containing monoamine oxidase (MAO) from Escherichia coli was overproduced in the periplasmic space by expression of the cloned gene. The purified MAO has been crystallized by means of the hanging drop technique using sodium citrate as a precipitant. The crystals belong to the orthorhombic system, space group P2(1)2(1)2(1), with unit cell dimensions of a = 136.1 A, b = 168.4 A and c = 81.6 A. The asymmetric unit contains one molecule of MAO, with a crystal volume per protein mass (Vm) of 2.88 A3/Da and a solvent content of 58% by volume. The crystals diffract X-rays to a resolution limit of at least 2.7 A and are resistant to X-ray radiation damage. They appear to be suitable for X-ray structure analysis.

摘要

通过克隆基因的表达，大肠杆菌含铜单胺氧化酶（MAO）在周质空间中过量产生。纯化后的MAO采用柠檬酸钠作为沉淀剂，通过悬滴法进行结晶。晶体属于正交晶系，空间群为P2(1)2(1)2(1)，晶胞参数为a = 136.1 Å，b = 168.4 Å，c = 81.6 Å。不对称单元包含一个MAO分子，每蛋白质质量的晶体体积（Vm）为2.88 Å³/Da，溶剂含量为58%（体积）。这些晶体的X射线衍射分辨率至少为2.7 Å，并且对X射线辐射损伤具有抗性。它们似乎适合进行X射线结构分析。