Identification of <i>Mycobacterium </i><i>t</i><i>uberculosis</i> H37Rv Integral Membrane Proteins by One-Dimensional Gel Electrophoresis and Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry Ying Xiong Michael J. Chalmers Fei Philip Gao Timothy A. Cross Alan G. Marshall 10.1021/pr0500049.s001 https://acs.figshare.com/articles/journal_contribution/Identification_of_i_Mycobacterium_i_i_t_i_i_uberculosis_i_H37Rv_Integral_Membrane_Proteins_by_One_Dimensional_Gel_Electrophoresis_and_Liquid_Chromatography_Electrospray_Ionization_Tandem_Mass_Spectrometry/3282469 Because many membrane-associated proteins represent potential drug targets, diagnostic probes, and components of vaccines, we have chosen to study the membrane proteins of <i>Mycobacterium tuberculosis</i> H37Rv. To remove cytosolic proteins and facilitate access to the integral membrane proteins, membrane fractions of <i>M. tuberculosis</i> 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 <i>M. tuberculosis</i> H37Rv proteins, including 42 integral membrane proteins, are described for the first time. Keywords: <i>Mycobacterium tuberculosis</i> H37Rv integral membrane protein • one-dimensional gel electrophoresis • membrane-associated proteins • nanoLC • ESI • MS/MS • protein identification • transmembrane α helix 2005-06-13 00:00:00 drug targets tuberculosis H 37Rv proteins MOWSE scores integral membrane proteins Mycobacterium t uberculosis H 37Rv Integral Membrane Proteins report 349 protein identifications 42 integral membrane proteins membrane proteins transmembrane α helix pH carbonate solution MS cytosolic proteins tryptic peptide Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry membrane fraction Mycobacterium tuberculosis H 37Rv 84 M 349 proteins signal sequence tuberculosis H 37Rv membrane fractions ESI 5 M urea