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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。

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1
Purification and crystallization of Escherichia coli pseudouridine synthase RluD.大肠杆菌假尿苷合酶RluD的纯化与结晶
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2
Crystal structure of the catalytic domain of RluD, the only rRNA pseudouridine synthase required for normal growth of Escherichia coli.RluD催化结构域的晶体结构,RluD是大肠杆菌正常生长所需的唯一rRNA假尿嘧啶合酶。
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Crystal structure of the RluD pseudouridine synthase catalytic module, an enzyme that modifies 23S rRNA and is essential for normal cell growth of Escherichia coli.RluD假尿苷合酶催化模块的晶体结构,该酶可修饰23S rRNA,对大肠杆菌的正常细胞生长至关重要。
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Isolation and properties of Escherichia coli 23S-RNA pseudouridine 1911, 1915, 1917 synthase (RluD).大肠杆菌23S - RNA假尿苷1911、1915、1917合成酶(RluD)的分离与特性
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A second function for pseudouridine synthases: A point mutant of RluD unable to form pseudouridines 1911, 1915, and 1917 in Escherichia coli 23S ribosomal RNA restores normal growth to an RluD-minus strain.假尿苷合酶的第二种功能:大肠杆菌23S核糖体RNA中无法形成假尿苷1911、1915和1917的RluD点突变体可使RluD缺失菌株恢复正常生长。
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引用本文的文献

1
RluD, a highly conserved pseudouridine synthase, modifies 50S subunits more specifically and efficiently than free 23S rRNA.RluD是一种高度保守的假尿苷合酶,它对50S亚基的修饰比游离的23S rRNA更具特异性和高效性。
RNA. 2007 Nov;13(11):1868-76. doi: 10.1261/rna.711207. Epub 2007 Sep 13.
2
Crystal structure of the catalytic domain of RluD, the only rRNA pseudouridine synthase required for normal growth of Escherichia coli.RluD催化结构域的晶体结构,RluD是大肠杆菌正常生长所需的唯一rRNA假尿嘧啶合酶。
RNA. 2004 Feb;10(2):231-9. doi: 10.1261/rna.5187404.