Characterization of the Salmonella typhimurium proteome by semi-automated two dimensional HPLC-mass spectrometry: detection of proteins implicated in multiple antibiotic resistance

Journal of Proteome Research
Nick G Coldham, Martin J Woodward

Abstract

The proteome of Salmonella enterica serovar Typhimurium was characterized by 2-dimensional HPLC mass spectrometry to provide a platform for subsequent proteomic investigations of low level multiple antibiotic resistance (MAR). Bacteria (2.15 +/- 0.23 x 10(10) cfu; mean +/- s.d.) were harvested from liquid culture and proteins differentially fractionated, on the basis of solubility, into preparations representative of the cytosol, cell envelope and outer membrane proteins (OMPs). These preparations were digested by treatment with trypsin and peptides separated into fractions (n = 20) by strong cation exchange chromatography (SCX). Tryptic peptides in each SCX fraction were further separated by reversed-phase chromatography and detected by mass spectrometry. Peptides were assigned to proteins and consensus rank listings compiled using SEQUEST. A total of 816 +/- 11 individual proteins were identified which included 371 +/- 33, 565 +/- 15 and 262 +/- 5 from the cytosolic, cell envelope and OMP preparations, respectively. A significant correlation was observed (r2 = 0.62 +/- 0.10; P < 0.0001) between consensus rank position for duplicate cell preparations and an average of 74 +/- 5% of proteins were common to both replicates. A tot...Continue Reading

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Citations

Apr 5, 2008·The Journal of Antimicrobial Chemotherapy·Mark A WebberLaura J V Piddock
Oct 13, 2009·Journal of Bacteriology·Sivagamisundaram ChavadiPaul R Wheeler
May 17, 2005·Expert Review of Proteomics·Celia G ZhangSandra L McCutchen-Maloney
May 17, 2005·Expert Review of Proteomics·Richard R DrakeStefan Gravenstein
Sep 10, 2014·International Journal of Molecular Sciences·Susana CorreiaGilberto Igrejas
Jun 15, 2013·Chemical Biology & Drug Design·Simona SutkuvienėRimantas Daugelavičius
Apr 13, 2010·Research in Microbiology·Kênia Valéria dos SantosLuiz de Macêdo Farias
Sep 10, 2008·Proteomics·Ansgar Poetsch, Dirk Wolters
Aug 29, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·E RehS Lamotte
Mar 19, 2015·Frontiers in Microbiology·Fiona FouhyFiona Walsh
May 27, 2016·Molecular BioSystems·Anita HorvatićMangesh Bhide
Jan 2, 2008·Journal of Proteome Research·Charles AnsongRichard D Smith
Oct 26, 2006·The Journal of Antimicrobial Chemotherapy·Nick G ColdhamMartin J Woodward
Jun 22, 2005·Expert Review of Proteomics·Andrew C PaolettiMichael P Washburn
May 11, 2006·Molecular & Cellular Proteomics : MCP·Joshua N AdkinsRichard D Smith
Jun 12, 2013·Metallomics : Integrated Biometal Science·Zhaojun YunGuibin Jiang
Nov 9, 2004·Journal of Mass Spectrometry : JMS
Jul 4, 2008·Journal of Proteome Research·Melkamu Getie-KebtieMichail A Alterman
Jul 26, 2007·Chemical Reviews·Marjorie L FournierMichael P Washburn

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