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