期刊
PROTEIN EXPRESSION AND PURIFICATION
卷 48, 期 1, 页码 49-55出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2006.01.015
关键词
O-antigen; Wzz; overexpression; secondary structure; interaction
类别
资金
- NIAID NIH HHS [R01 AI44040] Funding Source: Medline
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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