title: Probing of bismuth antiulcer drug targets in H. pylori by laser ablation-inductively coupled plasma mass spectrometry.
abstract: A method that allows partial denaturation of protein ligands in Bi- and Zn-protein complexes, leaving the metal coordination centre intact, was developed. It was based on the reduction of the S-S bridges with tris(2-carboxyl)phosphine followed by derivatization with iodoacetamide. Consequently conditions that allow the separation of Bi- and Zn-protein complexes using SDS electrophoresis were found. The separation efficiency was much higher than that in non-denaturating blue native electrophoresis. The method allowed the detection of seven Bi-binding protein candidates in H. pylori treated with bismuth subcitrate, some of which-fructose-bisphosphate aldolase (33.6 kDa), urease alpha subunit (26.4 kDa), and the 16.8 kDa proteins: 30S ribosomal protein S6 and neutrophil activating protein (NapA)-were bio-induced during the treatment. The method also allowed the monitoring of the changes in the Zn-proteome during treatment of H. pylori with the Bi-drug, which was found to increase the concentration of the Zn-binding proteins with particularly strong expression of the urease, S-adenosylmethionine synthetase and the above 16.8 kDa proteins.
authors.affiliation: Department of Chemistry, The University of Hong Kong, Hong Kong, PR China.
authors.forename: Cheuk-Nam
authors.initials: CN
authors.lastname: Tsang
authors.forename: Juliusz
authors.initials: J
authors.lastname: Bianga
authors.forename: Hongzhe
authors.initials: H
authors.lastname: Sun
authors.forename: Joanna
authors.initials: J
authors.lastname: Szpunar
authors.forename: Ryszard
authors.initials: R
authors.lastname: Lobinski
mesh_terms: D000897:Anti-Ulcer Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000378:metabolism; D001729:Bismuth / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D016480:Helicobacter pylori / Q000187:drug effects*; D007525:Isoelectric Focusing; D013058:Mass Spectrometry / Q000379:methods*; D014510:Urease / Q000378:metabolism
chemical_list: D000897:Anti-Ulcer Agents; D001426:Bacterial Proteins; D014510:Urease; D005634:Fructose-Bisphosphate Aldolase; D001729:Bismuth