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PMID:20149234

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Citation

Chooneea, D, Karlsson, R, Encheva, V, Arnold, C, Appleton, H and Shah, H (2010) Elucidation of the outer membrane proteome of Salmonella enterica serovar Typhimurium utilising a lipid-based protein immobilization technique. BMC Microbiol. 10:44

Abstract

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a major cause of human gastroenteritis worldwide. The outer membrane proteins expressed by S. Typhimurium mediate the process of adhesion and internalisation within the intestinal epithelium of the host thus influencing the progression of disease. Since the outer membrane proteins are surface-exposed, they provide attractive targets for the development of improved antimicrobial agents and vaccines. Various techniques have been developed for their characterisation, but issues such as carryover of cytosolic proteins still remain a problem. In this study we attempted to characterise the surface proteome of S. Typhimurium using Lipid-based Protein Immobilisation technology in the form of LPI FlowCells. No detergents are required and no sample clean up is needed prior to downstream analysis. The immobilised proteins can be digested with proteases in multiple steps to increase sequence coverage, and the peptides eluted can be characterised directly by liquid chromatography - tandem mass spectrometry (LC-MS/MS) and identified from mass spectral database searches.

Links

PubMed PMC2829538 Online version:10.1186/1471-2180-10-44

Keywords

Bacterial Outer Membrane Proteins/chemistry; Bacterial Outer Membrane Proteins/metabolism; Carbonates/chemistry; Cell Fractionation/methods; Chromatography, Liquid; Immobilized Proteins/chemistry; Immobilized Proteins/metabolism; Lipoproteins/chemistry; Lipoproteins/metabolism; Mass Spectrometry; Microscopy, Electron; Proteome/analysis; Proteomics/methods; Salmonella typhi/chemistry; Salmonella typhi/metabolism; Surface-Active Agents/chemistry

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