Global Analysis of the Membrane Subproteome of <i>Pseudomonas aeruginosa</i> Using Liquid Chromatography-Tandem Mass Spectrometry Josip Blonder Michael B. Goshe Wenzhong Xiao David G. Camp Mark Wingerd Ronald W. Davis Richard D. Smith 10.1021/pr034074w.s002 https://acs.figshare.com/articles/journal_contribution/Global_Analysis_of_the_Membrane_Subproteome_of_i_Pseudomonas_aeruginosa_i_Using_Liquid_Chromatography_Tandem_Mass_Spectrometry/3335863 <i>Pseudomonas aeruginosa</i> is one of the most significant opportunistic bacterial pathogens in humans causing infections and premature death in patients with cystic fibrosis, AIDS, severe burns, organ transplants, or cancer. Liquid chromatography coupled online with tandem mass spectrometry was used for the large-scale proteomic analysis of the <i>P. </i><i>aeruginosa</i> membrane subproteome. Concomitantly, an affinity labeling technique, using iodoacetyl-PEO biotin to tag cysteinyl-containing proteins, permitted the enrichment and detection of lower abundance membrane proteins. The application of these approaches resulted in the identification of 786 proteins. A total of 333 proteins (42%) had a minimum of one transmembrane domain (ranging from 1 to14) and 195 proteins were classified as hydrophobic based on their positive GRAVY values (ranging from 0.01 to 1.32). Key integral inner and outer membrane proteins involved in adaptation and antibiotic resistance were conclusively identified, including the detection of 53% of all predicted opr-type porins (outer integral membrane proteins) and all the components of the mexA-mexB-oprM transmembrane protein complex. This work represents one of the most comprehensive proteomic analyses of the membrane subproteome of <i>P. </i><i>aeruginosa</i> and for prokaryotes in general. Keywords: proteome • membrane proteins • low abundance • LC−MS/MS • affinity labeling 2004-06-14 00:00:00 integral membrane proteins 195 proteins tandem mass spectrometry Membrane Subproteome GRAVY values 786 proteins proteomic analysis abundance membrane proteins membrane proteins transmembrane domain aeruginosa membrane subproteome Liquid chromatography AIDS membrane subproteome LC proteomic analyses organ transplants Pseudomonas aeruginosa antibiotic resistance Global Analysis