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耐甲氧西林金黄色葡萄球菌Sar2676(一种泛酸合成酶)的纯化、结晶及数据收集

Purification, crystallization and data collection of methicillin-resistant Staphylococcus aureus Sar2676, a pantothenate synthetase.

作者信息

Seetharamappa Jaldappagari, Oke Muse, Liu Huanting, McMahon Stephen A, Johnson Kenneth A, Carter Lester, Dorward Mark, Zawadzki Michal, Overton Ian M, van Niekirk C A Johannes, Graham Shirley, Botting Catherine H, Taylor Garry L, White Malcolm F, Barton Geoffrey J, Coote Peter J, Naismith James H

机构信息

Scottish Structural Facility and Centre for Biomolecular Sciences, The University, St Andrews, Scotland, UK.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Jun 1;63(Pt 6):488-91. doi: 10.1107/S1744309107020362. Epub 2007 May 5.

Abstract

Sar2676, a pantothenate synthetase with a molecular weight of 31 419 Da from methicillin-resistant Staphylococcus aureus, has been expressed, purified and crystallized at 293 K. The protein crystallizes in a primitive triclinic lattice, with unit-cell parameters a = 45.3, b = 60.5, c = 117.6 A, alpha = 87.2, beta = 81.2, gamma = 68.4 degrees . A complete data set has been collected to 2.3 A resolution at the ESRF. Consideration of the likely solvent content suggested the asymmetric unit to contain four molecules. This has been confirmed by molecular-replacement phasing calculations, which give a solution with four monomers using a monomer of pantothenate synthetase from Escherichia coli (PDB code 1iho), which is 41% identical to Sar2676, as a search model.

摘要

Sar2676是一种来自耐甲氧西林金黄色葡萄球菌的泛酸合成酶,分子量为31419道尔顿,已在293K下进行表达、纯化和结晶。该蛋白质在原始三斜晶格中结晶,晶胞参数为a = 45.3、b = 60.5、c = 117.6 Å,α = 87.2、β = 81.2、γ = 68.4度。已在欧洲同步辐射装置(ESRF)上收集到分辨率为2.3 Å的完整数据集。对可能的溶剂含量的考虑表明不对称单元包含四个分子。这已通过分子置换相位计算得到证实,该计算使用来自大肠杆菌的泛酸合成酶单体(PDB代码1iho)作为搜索模型,得到了一个包含四个单体的解决方案,该大肠杆菌泛酸合成酶与Sar2676有41%的同源性。

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