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Identification of Mycobacterium tuberculosis H37Rv Integral Membrane Proteins by One-Dimensional Gel Electrophoresis and Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry
journal contribution
posted on 2005-06-13, 00:00 authored by Ying Xiong, Michael J. Chalmers, Fei Philip Gao, Timothy A. Cross, Alan G. MarshallBecause many membrane-associated proteins represent potential drug targets, diagnostic probes, and
components of vaccines, we have chosen to study the membrane proteins of Mycobacterium
tuberculosis H37Rv. To remove cytosolic proteins and facilitate access to the integral membrane
proteins, membrane fractions of M. tuberculosis H37Rv were intensely washed with 5 M urea and high
pH carbonate solution. One-dimensional SDS-PAGE, followed by enzymatic hydrolysis and nanoLC
electrospray ionization MS/MS, proved to be the most efficient way to identify the proteins contained
within the membrane fraction. Here we report 349 protein identifications in total, validated by at least
two tryptic peptide matches and MOWSE scores greater than 75. Of those 349 proteins, 100 are integral
membrane proteins with at least one predicted transmembrane α helix (excluding the possible signal
sequence). 84 M. tuberculosis H37Rv proteins, including 42 integral membrane proteins, are described
for the first time.
Keywords: Mycobacterium tuberculosis H37Rv integral membrane protein • one-dimensional gel electrophoresis
• membrane-associated proteins • nanoLC • ESI • MS/MS • protein identification • transmembrane α helix
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drug targetstuberculosis H 37Rv proteinsMOWSE scoresintegral membrane proteinsMycobacterium t uberculosis H 37Rv Integral Membrane Proteinsreport 349 protein identifications42 integral membrane proteinsmembrane proteinstransmembrane α helixpH carbonate solutionMScytosolic proteinstryptic peptideLiquid Chromatography Electrospray Ionization Tandem Mass Spectrometrymembrane fractionMycobacterium tuberculosis H 37Rv84 M349 proteinssignal sequencetuberculosis H 37Rvmembrane fractionsESI5 M urea
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