title: Characterization of Helicobacter pylori HP0231 (DsbK): role in disulfide bond formation, redox homeostasis and production of Helicobacter cystein-rich protein HcpE.
abstract: Helicobacter pylori is a human gastric pathogen that colonizes ∼ 50% of the world's population. It can cause gastritis, gastric or duodenal ulcers and also gastric cancer. The numerous side effects of available treatments and the emergence of antibiotic resistant strains are severe concerns that justify further research into H. pylori's pathogenic mechanisms. H. pylori produces secreted proteins that may play a role in virulence, including the Helicobacter cysteine-rich protein HcpE (aka HP0235). We demonstrate herein that HcpE is secreted in the culture supernatant both as a soluble protein and in association with outer membrane vesicles. We show that the structure of HcpE comprises an organized array of disulfide bonds. We identify DsbK (aka HP0231) as a folding factor necessary for HcpE production and secretion in H. pylori and show that recombinant DsbK can interact with and refold unprocessed, reduced HcpE in vitro. These experiments highlight the first biologically relevant substrate for DsbK. Furthermore, we show that DsbK has disulfide bond (Dsb) forming activity on reduced lysozyme and demonstrate a DsbA-type of activity for DsbK upon expression in E. coli, despite its similarity with DsbG. Finally, we show a role of DsbK in maintaining redox homeostasis in H. pylori.
authors.affiliation: Microbiology and Immunology, Western University, London, N6A5C1, Canada.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Lester
authors.forename: Sari
authors.initials: S
authors.lastname: Kichler
authors.forename: Brandon
authors.initials: B
authors.lastname: Oickle
authors.forename: Spencer
authors.initials: S
authors.lastname: Fairweather
authors.forename: Alexander
authors.initials: A
authors.lastname: Oberc
authors.forename: Jaspreet
authors.initials: J
authors.lastname: Chahal
authors.forename: Dinath
authors.initials: D
authors.lastname: Ratnayake
authors.forename: Carole
authors.initials: C
authors.lastname: Creuzenet
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016480:Helicobacter pylori / Q000378:metabolism*; D006706:Homeostasis; D008958:Models, Molecular; D009154:Mutation; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism; D017510:Protein Folding; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D004220:Disulfides; D010088:Oxidoreductases