4.3 Article

Comparison of surface proteomes of enterotoxigenic (ETEC) and commensal Escherichia coli strains

Journal

JOURNAL OF MICROBIOLOGICAL METHODS
Volume 83, Issue 1, Pages 13-19

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2010.07.011

Keywords

SILAC; Escherichia coli; Surface proteins; Vaccine; Proteomics

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Pathogenesis of enterotoxigenic Escherichia coli (ETEC) infections involves colonization of the small intestine mediated by cell-surface fimbriae (CS) or colonization fimbriae antigens (CFA). However, protection against reinfection of ETEC is also conferred by somatic antigens rather than by virulence factors. To discover ETEC specific somatic antigens, the surface proteome of the ETEC H10406 strain was compared with that of nonpathogenic E. coli K12 strains. In this study, we were using stable isotope labelling with amino acids in cell culture (SILAC) technology for the labelling and relative quantification of surface proteins in order to identify polypeptides that are specifically present on ETEC strains. Outer membrane proteins were isolated, separated by gel electrophoresis, and identified by mass spectrometry. Twenty-three differentially expressed cell-surface polypeptides of ETEC were identified and evaluated by bioinformatics for protein vaccine candidates. The combination of being surface-exposed and present differentially makes these polypeptides highly suitable as targets for antibodies and thus for use in passive or active immunisation/vaccination. (C) 2010 Elsevier B.V. All rights reserved.

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