大肠杆菌假尿苷合酶RluD的纯化与结晶

Purification and crystallization of Escherichia coli pseudouridine synthase RluD.

作者信息

Del Campo Mark, Ofengand James, Malhotra Arun

机构信息

Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, PO Box 016129, Miami, FL 33101-6129, USA.

出版信息

Acta Crystallogr D Biol Crystallogr. 2003 Oct;59(Pt 10):1871-3. doi: 10.1107/s0907444903018468. Epub 2003 Sep 19.

DOI:10.1107/s0907444903018468

PMID:14501142

Abstract

RluD is the pseudouridine (Psi) synthase responsible for forming Psi1911, Psi1915 and Psi1917 in Escherichia coli 23S RNA. Out of the 11 Psi synthases in E. coli, only cells lacking RluD show a severe growth defect. In addition, RluD belongs to the RluA family of Psi synthases, one of the two remaining families without a representative crystal structure. In this paper, the crystallization of selenomethionine-substituted RluD by the hanging-drop method is reported. The crystals diffract to 1.9 A and belong to space group P4(3)2(1)2, with unit-cell parameters a = b = 75.14, c = 181.81 A. Synchrotron radiation was used on a single crystal to collect a complete multiwavelength anomalous dispersion (MAD) data set to 2.0 A resolution.

摘要

RluD是负责在大肠杆菌23S RNA中形成假尿苷（Ψ）Psi1911、Psi1915和Psi1917的假尿苷合酶。在大肠杆菌的11种假尿苷合酶中，只有缺乏RluD的细胞表现出严重的生长缺陷。此外，RluD属于假尿苷合酶的RluA家族，是两个仍无代表性晶体结构的家族之一。本文报道了通过悬滴法对硒代甲硫氨酸取代的RluD进行结晶。晶体衍射至1.9 Å，属于空间群P4(3)2(1)2，晶胞参数a = b = 75.14，c = 181.81 Å。使用同步辐射对单晶进行照射，以收集分辨率为2.0 Å的完整多波长反常色散（MAD）数据集。