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A proteomic approach for the identification of bismuth-binding proteins in Helicobacter pylori

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Abstract

Helicobacter pylori is a major human pathogen that can cause peptic ulcers and chronic gastritis. Bismuth-based triple or quadruple therapies are commonly recommended for the treatment of H. pylori infections. However, the molecular mechanisms underlying treatment with bismuth are currently not fully understood. We have conducted a detailed comparative proteomic analysis of H. pylori cells both before and after treatment with colloidal bismuth subcitrate (CBS). Eight proteins were found to be significantly upregulated or downregulated in the presence of CBS (20 μg mL−1). Bismuth-induced oxidative stress was confirmed by detecting higher levels of lipid hydroperoxide (approximately 1.8 times) and hemin (approximately 3.4 times), in whole cell extracts of bismuth-treated H. pylori cells, compared with those from untreated cells. The presence of bismuth also led to an approximately eightfold decrease in cellular protease activities. Using immobilized-bismuth affinity chromatography, we isolated and subsequently identified seven bismuth-binding proteins from H. pylori cell extracts. The intracellular levels of four of these proteins (HspA, HspB, NapA and TsaA) were influenced by the addition of CBS, which strongly suggests that they interact directly with bismuth. The other bismuth-interacting proteins identified were two enzymes (fumarase and the urease subunit UreB), and a translational factor (Ef-Tu). Our data suggest that the inhibition of proteases, modulation of cellular oxidative stress and interference with nickel homeostasis may be key processes underlying the molecular mechanism of bismuth’s actions against H. pylori.

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Abbreviations

CBS:

Colloidal bismuth subcitrate

CHAPS:

3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate

2-DE:

Two-dimensional gel electrophoresis

DTT:

Dithiothreitol

FBS:

Fetal bovine serum

ICP:

Inductively coupled plasma

IEF:

Isoelectric focusing

IMAC:

Immobilized-metal affinity chromatography

LPO:

Lipid hydroperoxide

MALDI:

Matrix-assisted laser desorption/ionization

MS:

Mass spectrometry

NTA:

Nitrilotriacetate

PAGE:

Polyacylamide gel electrophoresis

pI:

Isoelectric point

PMSF:

Phenylmethylsulfonyl fluoride

ROS:

Reactive oxygen species

SDS:

Sodium dodecyl sulfate

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Acknowledgements

This work was supported by the Research Grants Council of Hong Kong (to H.S., HKU703904P and HKU704306P), the Research Fund for the Control of Infectious Diseases (to J.-D.H., 01030182), the Area of Excellence Scheme of the University Grants Committee, Institute of Molecular Technology for Drug Discovery and Synthesis) (AoE/P-10/01), Livzon Pharmaceutical Ltd and the University of Hong Kong. We are grateful to Jen-Fu Chiu (Department of Anatomy, Li Ka Shing Faculty of Medicine) for technical assistance.

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Authors and Affiliations

  1. Department of Chemistry and Open Laboratory of Chemical Biology, The University of Hong Kong, Pokfulam Road, Hong Kong, People’s Republic of China

    Ruiguang Ge, Xuesong Sun, Rory M. Watt, Qing-Yu He & Hongzhe Sun

  2. Department of Biochemistry, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, People’s Republic of China

    Julian A. Tanner & Jian-Dong Huang

  3. Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, People’s Republic of China

    Qing Gu, Benjamin Chun Yu Wong & Harry Huaxiang Xia

  4. Institute of Life and Health Engineering, Jinan University, Guangzhou, 510632, People’s Republic of China

    Qing-Yu He

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  1. Ruiguang Ge
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  2. Xuesong Sun
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Correspondence to Hongzhe Sun.

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Ge, R., Sun, X., Gu, Q. et al. A proteomic approach for the identification of bismuth-binding proteins in Helicobacter pylori . J Biol Inorg Chem 12, 831–842 (2007). https://doi.org/10.1007/s00775-007-0237-7

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  • DOI: https://doi.org/10.1007/s00775-007-0237-7

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