title: Overexpression and characterization of Wzz of Escherichia coli O86:H2.
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.
authors.affiliation: Department of Biochemistry and Chemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Hongjie
authors.initials: H
authors.lastname: Guo
authors.forename: Kaarina
authors.initials: K
authors.lastname: Lokko
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
authors.forename: Wen
authors.initials: W
authors.lastname: Yi
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Wu
authors.forename: Peng George
authors.initials: PG
authors.lastname: Wang
mesh_terms: D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003902:Detergents / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D005557:Formaldehyde / Q000737:chemistry / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005960:Glucosides / Q000378:metabolism; D008970:Molecular Weight; D019081:O Antigens / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D012995:Solubility
chemical_list: D003902:Detergents; D029968:Escherichia coli Proteins; D005960:Glucosides; D019081:O Antigens; D011993:Recombinant Fusion Proteins; C489586:wzzB protein, E coli; D005557:Formaldehyde; C040358:dodecyl maltoside