Kaarina Lokko - Overexpression and characterization of Wzz of Escherichia coli O86 : H2

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      Publication Details (including relevant citation   information):

      Protein Expression and   Purification 48(1):49-55, 2006

      DOI: 10.1016/j.pep.2006.01.015

      Abstract:

      O-Antigen plays a critical role in the bacterium-host interplay,   the chain length is an 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 O86:H2 in wzz mutant O86: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.

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