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PMID: 10777535 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Enterotoxigenic Escherichia coli secretes active heat-labile enterotoxin via outer membrane vesicles.

The Journal of biological chemistry ·Vol. 275 ·No. 17 ·2000-04-28 ·Pages 12489-96

Horstman AL, Kuehn MJ

Abstract

Escherichia coli and other Gram-negative bacteria produce outer membrane vesicles during normal growth. Vesicles may contribute to bacterial pathogenicity by serving as vehicles for toxins to encounter host cells. Enterotoxigenic E. coli (ETEC) vesicles were isolated from culture supernatants and purified on velocity gradients, thereby removing any soluble proteins and contaminants from the crude preparation. Vesicle protein profiles were similar but not identical to outer membranes and differed between strains. Most vesicle proteins were resistant to dissociation, suggesting they were integral or internal. Thin layer chromatography revealed that major outer membrane lipid components are present in vesicles. Cytoplasmic membranes and cytosol were absent in vesicles; however, alkaline phosphatase and AcrA, periplasmic residents, were localized to vesicles. In addition, physiologically active heat-labile enterotoxin (LT) was associated with ETEC vesicles. LT activity correlated directly with the gradient peak of vesicles, suggesting specific association, but could be removed from vesicles under dissociating conditions. Further analysis revealed that LT is enriched in vesicles and is located both inside and on the exterior of vesicles. The distinct protein composition of ETEC vesicles and their ability to carry toxin may contribute to the pathogenicity of ETEC strains.

MeSH Terms
Alkaline Phosphatase/metabolism Bacterial Toxins/biosynthesis Cell Fractionation Cell Membrane/metabolism Chromatography, Affinity Chromatography, Thin Layer Endopeptidases/metabolism Enterotoxins/biosynthesis Enzyme-Linked Immunosorbent Assay Escherichia coli/metabolism Escherichia coli Proteins Microscopy, Electron Temperature
Chemicals
Bacterial Toxins Enterotoxins Escherichia coli Proteins heat-labile enterotoxin, E coli Alkaline Phosphatase Endopeptidases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Horstman A L
Duke University Medical Center, Department of Biochemistry, Durham, North Carolina 27710, USA.
Kuehn M J
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.275.17.12489
Published
2000-04-28
Pages
12489-96
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC4347834
Subset
IM
Grants
NIAID NIH HHS · R21 AI063239 · United States
NIGMS NIH HHS · T32 GM007184 · United States
NIGMS NIH HHS · GM-07184 · United States
External Links
PubMed source DOI: 10.1074/jbc.275.17.12489 PMC Full Text View journal details
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