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PMID: 7608073 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Virulence factors are released from Pseudomonas aeruginosa in association with membrane vesicles during normal growth and exposure to gentamicin: a novel mechanism of enzyme secretion.

Journal of bacteriology ·Vol. 177 ·No. 14 ·1995-07-00 ·Pages 3998-4008

Kadurugamuwa JL, Beveridge TJ

Abstract

Pseudomonas aeruginosa blebs-off membrane vesicles (MVs) into culture medium during normal growth. Release of these vesicles increased approximately threefold after exposure of the organism to four times the MIC of gentamicin. Natural and gentamicin-induced membrane vesicles (n-MVs and g-MVs and g-MVs, respectively) were isolated by filtration and differential centrifugation, and several of their biological activities were characterized. Electron microscopy of both n-MVs and g-MVs revealed that they were spherical bilayer MVs with a diameter of 50 to 150 nm. Immunoelectron microscopy and Western blot (immunoblot) analysis of the vesicles demonstrated the presence of B-band lipopolysaccharide (LPS), with a slightly higher proportion of B-band LPS in g-MVs than in n-MVs. A-band LPS was occasionally detected in g-MVs but not in n-MVs. In addition to LPS, several enzymes, such as phospholipase C, protease, hemolysin, and alkaline phosphatase, which are known to contribute to the pathogenicity of Pseudomonas infections were found to be present in both vesicle types. Both types of vesicles contained DNA, with a significantly higher content in g-MVs. These vesicles could thus play an important role in genetic transformation and disease by serving as a transport vehicle for DNA and virulence factors and are presumably involved in septic shock.

MeSH Terms
Alkaline Phosphatase/metabolism Biological Transport Cell Membrane/drug effects,physiology,ultrastructure Electrophoresis, Polyacrylamide Gel Endopeptidases/metabolism Enzymes/metabolism Gentamicins/pharmacology Hemolysin Proteins/metabolism Lipopolysaccharides/chemistry,immunology,metabolism Microscopy, Immunoelectron Models, Biological Pseudomonas aeruginosa/drug effects,pathogenicity,physiology,ultrastructure Type C Phospholipases/metabolism Virulence
Chemicals
Enzymes Gentamicins Hemolysin Proteins Lipopolysaccharides Alkaline Phosphatase Type C Phospholipases Endopeptidases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kadurugamuwa J L
Department of Microbiology, College of Biological Science, University of Guelph, Ontario, Canada.
Beveridge T J
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.177.14.3998-4008.1995
Published
1995-07-00
Pages
3998-4008
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC177130
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.177.14.3998-4008.1995 PMC Full Text View journal details
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