Probing of bismuth antiulcer drug targets in H. pylori by laser ablation-inductively coupled plasma mass spectrometry

Metallomics. 2012 Mar;4(3):277-83. doi: 10.1039/c2mt00169a. Epub 2012 Jan 30.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Ulcer Agents / pharmacology*
  • Bacterial Proteins / metabolism
  • Bismuth / pharmacology*
  • Electrophoresis, Gel, Two-Dimensional
  • Fructose-Bisphosphate Aldolase / metabolism
  • Gene Expression Regulation, Bacterial / drug effects
  • Helicobacter pylori / drug effects*
  • Isoelectric Focusing
  • Mass Spectrometry / methods*
  • Urease / metabolism

Substances

  • Anti-Ulcer Agents
  • Bacterial Proteins
  • Urease
  • Fructose-Bisphosphate Aldolase
  • Bismuth