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. 1975 Mar;121(3):800–806. doi: 10.1128/jb.121.3.800-806.1975

Capacity of the outer membrane of a gram-negative marine bacterium in the presence of cations to prevent lysis by Triton X-100.

T Unemoto, R A MacLeod
PMCID: PMC246006  PMID: 1116995

Abstract

Cells of marine pseudomonad B-16 (ATCC 19855) washed with a solution containing 0.3 M NaCl, 50 mM MgCl2, and 10 mM KCl (complete salts) could be protected from lysis in a hypotonic environment if the suspending medium contained either 20 mM Mg2+, 40 mM Na+, or 300 mM K+. When the outer double-track layer (the outer membrane) of the cell envelope was removed to yield mureinoplasts, the Mg2+, Na+ or K+, requirements to prevent lysis were raised to 80, 210, and 400 mM, respectively. In the presence of 0.1% Triton X-100, 220, 320, and 360 mM Mg2+, Na+ or K+, respectively, prevented lysis of the normal cells. Mureinoplasts and protoplasts, however, lysed instantly in the presence of the detergent at all concentrations of Mg2+, Na+, or K+ tested up to 1.2 M. Thus, the structure of the outer membrane appears to be maintained by appropriate concentrations of Mg2+ or Na+ in a form preventing the penetration of Triton X-100 and thereby protecting the cytoplasmic membrane from dissolution by the detergent. K+ was effective in this capacity with cells washed with complete salts solution but not with cells washed with a solution of NaCl, suggesting that bound Mg2+ was required in the cell wall membrane for K+ to be effective in preventing lysis by the detergent. At high concentrations (1 M) K+ and Mg2+, but not Na+, appeared to destabilize the structure of the outer membrane in the presence of Triton X-100.

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Selected References

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  1. ABRAM D., GIBBONS N. E. The effect of chlorides of monovalent cations, urea, detergents, and heat on morphology and the turbidity of suspensions of red halophilic bacteria. Can J Microbiol. 1961 Oct;7:741–750. doi: 10.1139/m61-088. [DOI] [PubMed] [Google Scholar]
  2. Beacham I. R., Kahana R., Levy L., Yagil E. Mutants of Escherichia coli K-12 "cryptic," or deficient in 5'-nucleotidase (uridine diphosphate-sugar hydrolase) and 3'-nucleotidase (cyclic phosphodiesterase) activity. J Bacteriol. 1973 Nov;116(2):957–964. doi: 10.1128/jb.116.2.957-964.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buckmire F. L., MacLeod R. A. Evidence for the release at low salt concentration of a lipid-protein-carbohydrate complex from isolated envelopes and whole cells of a marine pseudomonad. Can J Microbiol. 1971 May;17(5):713–723. doi: 10.1139/m71-114. [DOI] [PubMed] [Google Scholar]
  4. Buckmire F. L., MacLeod R. A. Nutrition and metabolism of marine bacteria. XIV. On the mechanism of lysis of a marine bacterium. Can J Microbiol. 1965 Aug;11(4):677–691. doi: 10.1139/m65-091. [DOI] [PubMed] [Google Scholar]
  5. CHRISTIAN J. H., INGRAM M. Lysis of Vibrio costicolus by osmotic shock. J Gen Microbiol. 1959 Feb;20(1):32–42. doi: 10.1099/00221287-20-1-32. [DOI] [PubMed] [Google Scholar]
  6. De Voe I. W., Oginsky E. L. Antagonistic effect of monovalent cations in maintenance of cellular integrity of a marine bacterium. J Bacteriol. 1969 Jun;98(3):1355–1367. doi: 10.1128/jb.98.3.1355-1367.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DePamphilis M. L., Adler J. Attachment of flagellar basal bodies to the cell envelope: specific attachment to the outer, lipopolysaccharide membrane and the cyoplasmic membrane. J Bacteriol. 1971 Jan;105(1):396–407. doi: 10.1128/jb.105.1.396-407.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Forsberg C. W., Costerton J. W., Macleod R. A. Quantitation, chemical characteristics, and ultrastructure of the three outer cell wall layers of a gram-negative bacterium. J Bacteriol. 1970 Dec;104(3):1354–1368. doi: 10.1128/jb.104.3.1354-1368.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Forsberg C. W., Costerton J. W., Macleod R. A. Separation and localization of cell wall layers of a gram-negative bacterium. J Bacteriol. 1970 Dec;104(3):1338–1353. doi: 10.1128/jb.104.3.1338-1353.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gow J. A., DeVoe U. W., MacLeod R. A. Dissociation in a marine pseudomonad. Can J Microbiol. 1973 Jun;19(6):695–701. doi: 10.1139/m73-113. [DOI] [PubMed] [Google Scholar]
  11. Gustafsson P., Nordström K., Normark S. Outer penetration barrier of Escherichia coli K-12: kinetics of the uptake of gentian violet by wild type and envelope mutants. J Bacteriol. 1973 Nov;116(2):893–900. doi: 10.1128/jb.116.2.893-900.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lanyi J. K. Influence of electron transport on the interaction between membrane lipids and Triton X-100 in Halobacterium cutirubrum. Biochemistry. 1973 Mar 27;12(7):1433–1438. doi: 10.1021/bi00731a025. [DOI] [PubMed] [Google Scholar]
  14. Lanyi J. K. Salt-dependent properties of proteins from extremely halophilic bacteria. Bacteriol Rev. 1974 Sep;38(3):272–290. doi: 10.1128/br.38.3.272-290.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leive L., Shovlin V. K., Mergenhagen S. E. Physical, chemical, and immunological properties of lipopolysaccharide released from Escherichia coli by ethylenediaminetetraacetate. J Biol Chem. 1968 Dec 25;243(24):6384–6391. [PubMed] [Google Scholar]
  16. Martin E. L., MacLeod R. A. Isolation and chemical composition of the cytoplasmic membrane of a gram-negative bacterium. J Bacteriol. 1971 Mar;105(3):1160–1167. doi: 10.1128/jb.105.3.1160-1167.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Matula T. I., MacLeod R. A. Mechanism of optical effects in suspensions of a marine pseudomonad. J Bacteriol. 1969 Oct;100(1):403–410. doi: 10.1128/jb.100.1.403-410.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Matula T. I., Srivastava V. S., Wong P., MacLeod R. A. Transport and retention of K+ and other metabolites in a marine pseudomonad and their relation to the mechanism of optical effects. J Bacteriol. 1970 Jun;102(3):790–796. doi: 10.1128/jb.102.3.790-796.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  20. PETHICA B. A. Lysis by physical and chemical methods. J Gen Microbiol. 1958 Apr;18(2):473–480. doi: 10.1099/00221287-18-2-473. [DOI] [PubMed] [Google Scholar]
  21. Schnaitman C. A. Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. J Bacteriol. 1971 Oct;108(1):545–552. doi: 10.1128/jb.108.1.545-552.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sheu C. W., Freese E. Lipopolysaccharide layer protection of gram-negative bacteria against inhibition by long-chain fatty acids. J Bacteriol. 1973 Sep;115(3):869–875. doi: 10.1128/jb.115.3.869-875.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Thompson J., Costerton J. W., MacLeon R. A. K plus-dependent deplasmolysis of a marine pseudomonad plasmolyzed in a hypotonic solution. J Bacteriol. 1970 Jun;102(3):843–854. doi: 10.1128/jb.102.3.843-854.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thompson L. M., MacLeod R. A. Biochemical localization of alkaline phosphatase in the cell wall of a marine pseudomonad. J Bacteriol. 1974 Feb;117(2):819–825. doi: 10.1128/jb.117.2.819-825.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Unemoto T., Tsuruoka T., Hayashi M. Role of Na + and K + in preventing lysis of a slightly halophilic Vibrio alginolyticus. Can J Microbiol. 1973 May;19(5):563–571. doi: 10.1139/m73-093. [DOI] [PubMed] [Google Scholar]
  26. Wolf-Watz H., Normark S., Bloom G. D. Rapid method for isolation of large quantities of outer membrane from Escherichia coli K-12 and its application to the study of envelope mutants. J Bacteriol. 1973 Sep;115(3):1191–1197. doi: 10.1128/jb.115.3.1191-1197.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]

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