pr034074w_si_002.pdf (155.75 kB)
Global Analysis of the Membrane Subproteome of Pseudomonas aeruginosa Using Liquid Chromatography-Tandem Mass Spectrometry
journal contribution
posted on 2004-06-14, 00:00 authored by Josip Blonder, Michael B. Goshe, Wenzhong Xiao, David G. Camp, Mark Wingerd, Ronald W. Davis, Richard D. SmithPseudomonas aeruginosa 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 P. aeruginosa 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 P. aeruginosa
and for prokaryotes in general.
Keywords: proteome • membrane proteins • low abundance • LC−MS/MS • affinity labeling
History
Usage metrics
Categories
Keywords
integral membrane proteins195 proteinstandem mass spectrometryMembrane SubproteomeGRAVY values786 proteinsproteomic analysisabundance membrane proteinsmembrane proteinstransmembrane domainaeruginosa membrane subproteomeLiquid chromatographyAIDSmembrane subproteomeLCproteomic analysesorgan transplantsPseudomonas aeruginosaantibiotic resistanceGlobal Analysis
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC