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. 1996 May;178(10):2767–2774. doi: 10.1128/jb.178.10.2767-2774.1996

Bacteriolytic effect of membrane vesicles from Pseudomonas aeruginosa on other bacteria including pathogens: conceptually new antibiotics.

J L Kadurugamuwa 1, T J Beveridge 1
PMCID: PMC178010  PMID: 8631663

Abstract

Pseudomonas aeruginosa releases membrane vesicles (MVs) filled with periplasmic components during normal growth, and the quantity of these vesicles can be increased by brief exposure to gentamicin. Natural and gentamicin-induced membrane vesicles (n-MVs and g-MVs, respectively) are subtly different from one another, but both contain several important virulence factors, including hydrolytic enzyme factors (J. L. Kadurugamuwa and T. J. Beveridge, J. Bacteriol. 177:3998-4008, 1995). Peptidoglycan hydrolases (autolysins) were detected in both MV types, especially a periplasmic 26-kDa autolysin whose expression has been related to growth phase (Z. Li, A. J. Clarke, and T. J. Beveridge, J. Bacteriol. 178:2479-2488, 1996). g-MVs possessed slightly higher autolysin activity and, at the same time, small quantities of gentamicin. Both MV types hydrolyzed isolated gram-positive and gram-negative murein sacculi and were also capable of hydrolyzing several glycyl peptides. Because the MVs were bilayered, they readily fused with the outer membrane of gram-negative bacteria. They also adhered to the cell wall of gram-positive bacteria. g-MVs were more effective in lysing other bacteria because, in addition to the autolysins, they also contained small amounts of gentamicin. The bactericidal activity was 2.5 times the MIC of gentamicin, which demonstrates the synergistic effect of the antibiotic with the autolysins. n-MVs were capable of killing cultures of P. aeruginosa with permeability resistance against gentamicin, indicating that the fusion of n-MV to the outer membrane liberated autolysins into the periplasm, where they degraded the peptidoglycan and lysed the cells. g-MVs had even greater killing power since they liberated both gentamicin and autolysins into these resistant cells. These findings may help develop a conceptually new group of antibiotics designed to be effective against hard-to-kill bacteria.

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Selected References

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  1. Angus B. L., Fyfe J. A., Hancock R. E. Mapping and characterization of two mutations to antibiotic supersusceptibility in Pseudomonas aeruginosa. J Gen Microbiol. 1987 Oct;133(10):2905–2914. doi: 10.1099/00221287-133-10-2905. [DOI] [PubMed] [Google Scholar]
  2. Bernadsky G., Beveridge T. J., Clarke A. J. Analysis of the sodium dodecyl sulfate-stable peptidoglycan autolysins of select gram-negative pathogens by using renaturing polyacrylamide gel electrophoresis. J Bacteriol. 1994 Sep;176(17):5225–5232. doi: 10.1128/jb.176.17.5225-5232.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beveridge T. J. The bacterial surface: general considerations towards design and function. Can J Microbiol. 1988 Apr;34(4):363–372. doi: 10.1139/m88-067. [DOI] [PubMed] [Google Scholar]
  4. Beveridge T. J. Ultrastructure, chemistry, and function of the bacterial wall. Int Rev Cytol. 1981;72:229–317. doi: 10.1016/s0074-7696(08)61198-5. [DOI] [PubMed] [Google Scholar]
  5. Bierbaum G., Sahl H. G. Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase. J Bacteriol. 1987 Dec;169(12):5452–5458. doi: 10.1128/jb.169.12.5452-5458.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brito N., Falcón M. A., Carnicero A., Gutiérrez-Navarro A. M., Mansito T. B. Purification and peptidase activity of a bacteriolytic extracellular enzyme from Pseudomonas aeruginosa. Res Microbiol. 1989 Feb;140(2):125–137. doi: 10.1016/0923-2508(89)90046-6. [DOI] [PubMed] [Google Scholar]
  7. Bryan L. E., O'Hara K., Wong S. Lipopolysaccharide changes in impermeability-type aminoglycoside resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1984 Aug;26(2):250–255. doi: 10.1128/aac.26.2.250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burke M. E., Pattee P. A. Purification and characterization of a staphylolytic enzyme from Pseudomonas aeruginosa. J Bacteriol. 1967 Mar;93(3):860–865. doi: 10.1128/jb.93.3.860-865.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chatterjee S. N., Das J. Electron microscopic observations on the excretion of cell-wall material by Vibrio cholerae. J Gen Microbiol. 1967 Oct;49(1):1–11. doi: 10.1099/00221287-49-1-1. [DOI] [PubMed] [Google Scholar]
  10. Cozens R. M., Markiewicz Z., Tuomanen E. Role of autolysins in the activities of imipenem and CGP 31608, a novel penem, against slowly growing bacteria. Antimicrob Agents Chemother. 1989 Oct;33(10):1819–1821. doi: 10.1128/aac.33.10.1819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Devoe I. W., Gilchrist J. E. Release of endotoxin in the form of cell wall blebs during in vitro growth of Neisseria meningitidis. J Exp Med. 1973 Nov 1;138(5):1156–1167. doi: 10.1084/jem.138.5.1156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Filloux A., Bally M., Ball G., Akrim M., Tommassen J., Lazdunski A. Protein secretion in gram-negative bacteria: transport across the outer membrane involves common mechanisms in different bacteria. EMBO J. 1990 Dec;9(13):4323–4329. doi: 10.1002/j.1460-2075.1990.tb07881.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Glauner B., Höltje J. V., Schwarz U. The composition of the murein of Escherichia coli. J Biol Chem. 1988 Jul 25;263(21):10088–10095. [PubMed] [Google Scholar]
  14. Höltje J. V., Tuomanen E. I. The murein hydrolases of Escherichia coli: properties, functions and impact on the course of infections in vivo. J Gen Microbiol. 1991 Mar;137(3):441–454. doi: 10.1099/00221287-137-3-441. [DOI] [PubMed] [Google Scholar]
  15. Kadurugamuwa J. L., Beveridge T. J. 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. J Bacteriol. 1995 Jul;177(14):3998–4008. doi: 10.1128/jb.177.14.3998-4008.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kadurugamuwa J. L., Clarke A. J., Beveridge T. J. Surface action of gentamicin on Pseudomonas aeruginosa. J Bacteriol. 1993 Sep;175(18):5798–5805. doi: 10.1128/jb.175.18.5798-5805.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kadurugamuwa J. L., Lam J. S., Beveridge T. J. Interaction of gentamicin with the A band and B band lipopolysaccharides of Pseudomonas aeruginosa and its possible lethal effect. Antimicrob Agents Chemother. 1993 Apr;37(4):715–721. doi: 10.1128/aac.37.4.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kessler E., Safrin M., Olson J. C., Ohman D. E. Secreted LasA of Pseudomonas aeruginosa is a staphylolytic protease. J Biol Chem. 1993 Apr 5;268(10):7503–7508. [PubMed] [Google Scholar]
  19. Knirel YuA, Vinogradov E. V., Kocharova N. A., Paramonov N. A., Kochetkov N. K., Dmitriev B. A., Stanislavsky E. S., Lányi B. The structure of O-specific polysaccharides and serological classification of Pseudomonas aeruginosa (a review). Acta Microbiol Hung. 1988;35(1):3–24. [PubMed] [Google Scholar]
  20. Koch A. L. Additional arguments for the key role of "smart" autolysins in the enlargement of the wall of gram-negative bacteria. Res Microbiol. 1990 Jun;141(5):529–541. doi: 10.1016/0923-2508(90)90017-k. [DOI] [PubMed] [Google Scholar]
  21. Kondo K., Takade A., Amako K. Release of the outer membrane vesicles from Vibrio cholerae and Vibrio parahaemolyticus. Microbiol Immunol. 1993;37(2):149–152. doi: 10.1111/j.1348-0421.1993.tb03192.x. [DOI] [PubMed] [Google Scholar]
  22. Lache M., Hearn W. R., Zyskind J. W., Tipper D. J., Strominger J. L. Specificity of a bacteriolytic enzyme from Pseudomonas aeruginosa. J Bacteriol. 1969 Oct;100(1):254–259. doi: 10.1128/jb.100.1.254-259.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lam J. S., Graham L. L., Lightfoot J., Dasgupta T., Beveridge T. J. Ultrastructural examination of the lipopolysaccharides of Pseudomonas aeruginosa strains and their isogenic rough mutants by freeze-substitution. J Bacteriol. 1992 Nov;174(22):7159–7167. doi: 10.1128/jb.174.22.7159-7167.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lazdunski A., Guzzo J., Filloux A., Bally M., Murgier M. Secretion of extracellular proteins by Pseudomonas aeruginosa. Biochimie. 1990 Feb-Mar;72(2-3):147–156. doi: 10.1016/0300-9084(90)90140-c. [DOI] [PubMed] [Google Scholar]
  25. Li Z., Clarke A. J., Beveridge T. J. A major autolysin of Pseudomonas aeruginosa: subcellular distribution, potential role in cell growth and division and secretion in surface membrane vesicles. J Bacteriol. 1996 May;178(9):2479–2488. doi: 10.1128/jb.178.9.2479-2488.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lory S. Determinants of extracellular protein secretion in gram-negative bacteria. J Bacteriol. 1992 Jun;174(11):3423–3428. doi: 10.1128/jb.174.11.3423-3428.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mansito T. B., Falcón M. A., Moreno J., Carnicero A., Gutierrez-Navarro A. M. Effects of staphylolytic enzymes from Pseudomonas aeruginosa on the growth and ultrastructure of Staphylococcus aureus. Microbios. 1987;49(198):55–64. [PubMed] [Google Scholar]
  28. Martin N. L., Beveridge T. J. Gentamicin interaction with Pseudomonas aeruginosa cell envelope. Antimicrob Agents Chemother. 1986 Jun;29(6):1079–1087. doi: 10.1128/aac.29.6.1079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mayrand D., Holt S. C. Biology of asaccharolytic black-pigmented Bacteroides species. Microbiol Rev. 1988 Mar;52(1):134–152. doi: 10.1128/mr.52.1.134-152.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Park S., Galloway D. R. Purification and characterization of LasD: a second staphylolytic proteinase produced by Pseudomonas aeruginosa. Mol Microbiol. 1995 Apr;16(2):263–270. doi: 10.1111/j.1365-2958.1995.tb02298.x. [DOI] [PubMed] [Google Scholar]
  31. Parr T. R., Jr, Bayer A. S. Mechanisms of aminoglycoside resistance in variants of Pseudomonas aeruginosa isolated during treatment of experimental endocarditis in rabbits. J Infect Dis. 1988 Nov;158(5):1003–1010. doi: 10.1093/infdis/158.5.1003. [DOI] [PubMed] [Google Scholar]
  32. Pugsley A. P. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. doi: 10.1128/mr.57.1.50-108.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rivera M., Bryan L. E., Hancock R. E., McGroarty E. J. Heterogeneity of lipopolysaccharides from Pseudomonas aeruginosa: analysis of lipopolysaccharide chain length. J Bacteriol. 1988 Feb;170(2):512–521. doi: 10.1128/jb.170.2.512-521.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schleifer K. H., Kandler O. Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev. 1972 Dec;36(4):407–477. doi: 10.1128/br.36.4.407-477.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Strominger J. L., Ghuysen J. M. Mechanisms of enzymatic bacteriaolysis. Cell walls of bacteri are solubilized by action of either specific carbohydrases or specific peptidases. Science. 1967 Apr 14;156(3772):213–221. doi: 10.1126/science.156.3772.213. [DOI] [PubMed] [Google Scholar]
  36. Szabó I., Penyige A., Barabás G., Barabás J. Effect of aminoglycoside antibiotics on the autolytic enzyme of Streptomyces griseus. Arch Microbiol. 1990;155(1):99–102. doi: 10.1007/BF00291282. [DOI] [PubMed] [Google Scholar]
  37. Verwer R. W., Nanninga N., Keck W., Schwarz U. Arrangement of glycan chains in the sacculus of Escherichia coli. J Bacteriol. 1978 Nov;136(2):723–729. doi: 10.1128/jb.136.2.723-729.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wai S. N., Takade A., Amako K. The release of outer membrane vesicles from the strains of enterotoxigenic Escherichia coli. Microbiol Immunol. 1995;39(7):451–456. doi: 10.1111/j.1348-0421.1995.tb02228.x. [DOI] [PubMed] [Google Scholar]
  39. Walker S. G., Beveridge T. J. Amikacin disrupts the cell envelope of Pseudomonas aeruginosa ATCC 9027. Can J Microbiol. 1988 Jan;34(1):12–18. doi: 10.1139/m88-003. [DOI] [PubMed] [Google Scholar]
  40. Wandersman C. Secretion across the bacterial outer membrane. Trends Genet. 1992 Sep;8(9):317–322. doi: 10.1016/0168-9525(92)90264-5. [DOI] [PubMed] [Google Scholar]
  41. Watt S. R., Clarke A. J. Initial characterization of two extracellular autolysins from Pseudomonas aeruginosa PAO1. J Bacteriol. 1994 Aug;176(15):4784–4789. doi: 10.1128/jb.176.15.4784-4789.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Watt S. R., Clarke A. J. Role of autolysins in the EDTA-induced lysis of Pseudomonas aeruginosa. FEMS Microbiol Lett. 1994 Nov 15;124(1):113–119. doi: 10.1111/j.1574-6968.1994.tb07270.x. [DOI] [PubMed] [Google Scholar]
  43. Whitmire W. M., Garon C. F. Specific and nonspecific responses of murine B cells to membrane blebs of Borrelia burgdorferi. Infect Immun. 1993 Apr;61(4):1460–1467. doi: 10.1128/iai.61.4.1460-1467.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wilkinson S. G. Composition and structure of lipopolysaccharides from Pseudomonas aeruginosa. Rev Infect Dis. 1983 Nov-Dec;5 (Suppl 5):S941–S949. doi: 10.1093/clinids/5.supplement_5.s941. [DOI] [PubMed] [Google Scholar]
  45. Wispelwey B., Hansen E. J., Scheld W. M. Haemophilus influenzae outer membrane vesicle-induced blood-brain barrier permeability during experimental meningitis. Infect Immun. 1989 Aug;57(8):2559–2562. doi: 10.1128/iai.57.8.2559-2562.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wretlind B., Pavlovskis O. R. Pseudomonas aeruginosa elastase and its role in pseudomonas infections. Rev Infect Dis. 1983 Nov-Dec;5 (Suppl 5):S998–1004. doi: 10.1093/clinids/5.supplement_5.s998. [DOI] [PubMed] [Google Scholar]

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