Comparative Proteomics Analysis of Sarcosine Insoluble Outer Membrane Proteins from Clarithromycin Resistant and Sensitive Strains of Helicobacter pylori

Rebecca Smiley; James Bailey; Mahadevan Sethuraman; Norberto Posecion; M. Showkat Ali

doi:10.1007/s12275-013-3029-5

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Comparative Proteomics Analysis of Sarcosine Insoluble Outer Membrane Proteins from Clarithromycin Resistant and Sensitive Strains of Helicobacter pylori: Rebecca Smiley ¹, James Bailey ², Mahadevan Sethuraman ², Norberto Posecion ², M. Showkat Ali ^1,2; Journal of Microbiology 2013;51(5):612-618.
DOI: https://doi.org/10.1007/s12275-013-3029-5
Published online: October 31, 2013

¹Department of Clinical Investigation, William Beaumont Army Medical Center, 5005 Piedras Street, El Paso, TX 79920-5001, USA, ²Texas Tech Health Science Center Paul L. Foster School of Medicine, 5001 El Paso Drive, El Paso, TX 79905-2709, USA¹Department of Clinical Investigation, William Beaumont Army Medical Center, 5005 Piedras Street, El Paso, TX 79920-5001, USA, ²Texas Tech Health Science Center Paul L. Foster School of Medicine, 5001 El Paso Drive, El Paso, TX 79905-2709, USA

Corresponding author: M. Showkat Ali , Tel: +1-915-742-6041,

Received: 14 January 2013 • Accepted: 25 April 2013

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Abstract

Helicobacter pylori causes disease manifestations in humans including chronic gastric and peptic ulcers, gastric cancer, and lymphoid tissue lymphoma. Increasing rates of H. pylori clarithromycin resistance has led to higher rates of disease development. Because antibiotic resistance involves modifications of outer membrane proteins (OMP) in other Gram-negative bacteria, this study focuses on identification of H. pylori OMP’s using comparative proteomic analyses of clarithromycin-susceptible and -resistant H. pylori strains. Comparative proteomics analyses of isolated sarcosine-insoluble OMP fractions from clarithromycin-susceptible and -resistant H. pylori strains were performed by 1) one dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis protein separation and 2) in-gel digestion of the isolated proteins and mass spectrometry analysis by Matrix Assisted Laser Desorption Ionization-tandem mass spectrometry. Iron-regulated membrane protein, UreaseB, EF-Tu, and putative OMP were down-regulated; HopT (BabB) transmembrane protein, HofC, and OMP31 were up-regulated in clarithromycin-resistant H. pylori. Western blotting and real time PCR, respectively, validated UreaseB subunit and EF-Tu changes at the protein level, and mRNA expression of HofC and HopT. This limited proteomic study provides evidence that alteration of the outer membrane proteins’ profile may be a novel mechanism involved in clarithromycin resistance in H. pylori.

Keywords: outer membrane protein;clarithromycin;antibiotic resistance

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Comparative Proteomics Analysis of Sarcosine Insoluble Outer Membrane Proteins from Clarithromycin Resistant and Sensitive Strains of Helicobacter pylori

J. Microbiol. 2013;51(5):612-618. Published online October 31, 2013

DOI: https://doi.org/10.1007/s12275-013-3029-5

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