Overexpression and characterization of Wzz of Escherichia coli O86:H2

被引：30
|
作者：
Guo, Hongjie [1 ]
Lokko, Kaarina [1 ]
Zhang, Yun [1 ]
Yi, Wen [1 ]
Wu, Zhengrong [1 ]
Wang, Peng George [1 ]
机构：
[1] Ohio State Univ, Dept Biochem & Chem, Columbus, OH 43210 USA
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2006年 / 48卷 / 01期
关键词：
O-antigen; Wzz; overexpression; secondary structure; interaction;
D O I：
10.1016/j.pep.2006.01.015
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
O-Antigen plays a critical role in the bacterium-host interplay. the chain length is in important factor in O-antigen functions. Wzz protein is responsible for O-antigen chain length regulation, but the mechanism is still unknown. Here, we overexpressed the Wzz of Escherichia coli 086:H2 in wzz mutant 086:H2 strain, the yield can achieve 15 mg/L. The recombinant Wzz was purified to 99% purity in dodecylmaltoside by sequential Ni-affinity chromatography and anion-exchange. Size exclusion chromatography and in vivo cross-linking experiments both showed that Wzz formed tetramer. Furthermore. analysis with Circular dichroism revealed that the predominant structural composition in Wzz is alpha-helices, and incubation with O-antigen significantly changed Wzz conformation. The results suggested that Wzz protein can interact with O-antigen. (c) 2006 Elsevier Inc. All rights reserved.
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页码：49 / 55
页数：7
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