Crystallization and preliminary X-ray diffraction analysis of the lytic transglycosylase MltE from Escherichia coli

被引:7
作者
Artola-Recolons, Cecilia [1 ]
Llarrull, Leticia I. [2 ]
Lastochkin, Elena [2 ]
Mobashery, Shahriar [2 ]
Hermoso, Juan A. [1 ]
机构
[1] Inst Quim Fis, Consejo Super Invest Cient, Dept Crystallog & Struct Biol, Madrid 28006, Spain
[2] Univ Notre Dame, Dept Chem & Biochem, Notre Dame, IN 46556 USA
来源
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | 2011年 / 67卷
基金
美国国家卫生研究院;
关键词
CRYSTAL-STRUCTURES;
D O I
10.1107/S1744309110049171
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
MltE from Escherichia coli (193 amino acids, 21 380 Da) is a lytic transglycosylase that initiates the first step of cell-wall recycling. This enzyme is responsible for the cleavage of the cell-wall peptidoglycan at the beta-1,4-glycosidic bond between the N-acetylglucosamine and N-acetylmuramic acid units. At the end this reaction generates a disaccharide that is internalized and initiates the recycling process. To obtain insights into the biological functions of MltE, crystallization trials were performed and crystals of MltE protein that were suitable for X-ray diffraction analysis were obtained. The MltE protein of E. coli was crystallized using the hanging-drop vapour-diffusion method at 291 K. Crystals grew from a mixture consisting of 28% polyethylene glycol 4000, 0.1 M Tris pH 8.4 and 0.2 M magnesium chloride. Further optimization was performed using the microbatch technique. Single crystals were obtained that belonged to the orthorhombic space group C222(1), with unit-cell parameters a = 123.32, b = 183.93, c = 35.29 angstrom, and diffracted to a resolution of 2.1 angstrom.
引用
收藏
页码:161 / 163
页数:3
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