Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1975 Jul;8(1):95–104. doi: 10.1128/aac.8.1.95

Action of Polymyxin B on Bacterial Membranes: Morphological Changes in the Cytoplasm and in the Outer Membrane of Salmonella typhimurium and Escherichia coli B

Peter R G Schindler 1, Michael Teuber 1
PMCID: PMC429267  PMID: 169730

Abstract

Though the primary action of the cationic antibiotic polymyxin B is against the membrane of susceptible bacteria, severe morphological changes are detected in the cytoplasm. Using fluorescence microscopy and a mono-N-dansyl-polymyxin B derivative, we could demonstrate aggregations of the antibiotic with cellular material, possibly nucleic acids and/or ribosomes. These aggregations were only produced by minimum inhibitory or higher concentrations of the antibiotic as shown with Salmonella and Escherichia strains differing in their polymyxin susceptibility. The outer membrane of Salmonella typhimurium revealed characteristic blebs when treated with polymyxin B. This was investigated by the gentle methods of spray-freezing and freeze-etching. The obtained electron micrographs suggest that the polymyxin-induced blebs are projections of the outer monolayer of the outer membrane. A possible mechanism of penetration of polymyxin B through the cell envelope of gram-negative bacteria is presented.

Full text

PDF
100

Images in this article

97Image
on p.97
99Image
on p.99
100Image
on p.100
101Image
on p.101
101Image
on p.101
102Image
on p.102

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bachmann L., Schmitt W. W. Improved cryofixation applicable to freeze etching. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2149–2152. doi: 10.1073/pnas.68.9.2149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bader J., Teuber M. Action of polymyxin B on bacterial membranes. 1. Binding to the O-antigenic lipopolysaccharide of Salmonella typhimurium. Z Naturforsch C. 1973 Jul-Aug;28(7):422–430. [PubMed] [Google Scholar]
  3. Bayer M. E., Remsen C. C. Structure of Escherichia coli after freeze-etching. J Bacteriol. 1970 Jan;101(1):304–313. doi: 10.1128/jb.101.1.304-313.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bayer M. E. Ultrastructure and organization of the bacterial envelope. Ann N Y Acad Sci. 1974 May 10;235(0):6–28. doi: 10.1111/j.1749-6632.1974.tb43254.x. [DOI] [PubMed] [Google Scholar]
  5. Cerny G., Teuber M. Differential release of periplasmic versus cytoplasmic enzymes from Escherichia coli B by polymixin B. Arch Mikrobiol. 1971;78(2):166–179. doi: 10.1007/BF00424873. [DOI] [PubMed] [Google Scholar]
  6. FEW A. V., SCHULMAN J. H. The absorption of polymyxin E by bacteria and bacterial cell walls and its bactericidal action. J Gen Microbiol. 1953 Dec;9(3):454–466. doi: 10.1099/00221287-9-3-454. [DOI] [PubMed] [Google Scholar]
  7. Glauert A. M., Thornley M. J. The topography of the bacterial cell wall. Annu Rev Microbiol. 1969;23:159–198. doi: 10.1146/annurev.mi.23.100169.001111. [DOI] [PubMed] [Google Scholar]
  8. Handley P. S., Quesnel L. B., Sturgiss M. M. Ultrastructural changes produced in Proteus vulgaris by a synergistic combination of colistin and sulphadiazine. Microbios. 1974 May;10(40):211–223. [PubMed] [Google Scholar]
  9. HsuChen C. C., Feingold D. S. The mechanism of polymyxin B action and selectivity toward biologic membranes. Biochemistry. 1973 May 22;12(11):2105–2111. doi: 10.1021/bi00735a014. [DOI] [PubMed] [Google Scholar]
  10. Imai M., Inoue K., Nojima S. Effect of polymyxin B on liposomal membranes derived from Escherichia coli lipids. Biochim Biophys Acta. 1975 Jan 14;375(1):130–137. doi: 10.1016/0005-2736(75)90078-4. [DOI] [PubMed] [Google Scholar]
  11. Koike M., Iida K. Effect of polymyxin on the bacteriophage receptors of the cell walls of gram-negative bacteria. J Bacteriol. 1971 Dec;108(3):1402–1411. doi: 10.1128/jb.108.3.1402-1411.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Koike M., Iida K., Matsuo T. Electron microscopic studies on mode of action of polymyxin. J Bacteriol. 1969 Jan;97(1):448–452. doi: 10.1128/jb.97.1.448-452.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leive L. The barrier function of the gram-negative envelope. Ann N Y Acad Sci. 1974 May 10;235(0):109–129. doi: 10.1111/j.1749-6632.1974.tb43261.x. [DOI] [PubMed] [Google Scholar]
  14. Mizushima S., Yamada H. Isolation and characterization of two outer membrane preparations from Escherichia coli. Biochim Biophys Acta. 1975 Jan 14;375(1):44–53. doi: 10.1016/0005-2736(75)90071-1. [DOI] [PubMed] [Google Scholar]
  15. NEWTON B. A. The properties and mode of action of the polymyxins. Bacteriol Rev. 1956 Mar;20(1):14–27. doi: 10.1128/br.20.1.14-27.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nakajima K., Kawamata J. Studies on the mechanism of action of colistin. I. Formation of an insoluble complex with nucleic acids. Biken J. 1965 Dec;8(4):225–231. [PubMed] [Google Scholar]
  17. Nanninga N. Ultrastructure of the cell envelope of Escherichia coli B after freeze-etching. J Bacteriol. 1970 Jan;101(1):297–303. doi: 10.1128/jb.101.1.297-303.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Payne J. W., Gilvarg C. Size restriction on peptide utilization in Escherichia coli. J Biol Chem. 1968 Dec 10;243(23):6291–6299. [PubMed] [Google Scholar]
  20. Pontefract R. D., Bergeron G., Thatcher F. S. Mesosomes in Escherichia coli. J Bacteriol. 1969 Jan;97(1):367–375. doi: 10.1128/jb.97.1.367-375.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rifkind D. Studies on the interaction between endotoxin and polymyxin B. J Infect Dis. 1967 Dec;117(5):433–438. doi: 10.1093/infdis/117.5.433. [DOI] [PubMed] [Google Scholar]
  22. Ryter A. Association of the nucleus and the membrane of bacteria: a morphological study. Bacteriol Rev. 1968 Mar;32(1):39–54. doi: 10.1128/br.32.1.39-54.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sheetz M. P., Singer S. J. Biological membranes as bilayer couples. A molecular mechanism of drug-erythrocyte interactions. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4457–4461. doi: 10.1073/pnas.71.11.4457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Singh A. P., Cheng K. J., Costerton J. W., Idziak E. S., Ingram J. M. Sensitivity of normal and mutant strains of Escherichia coli to actinomycin-D. Can J Microbiol. 1972 Jun;18(6):909–915. doi: 10.1139/m72-139. [DOI] [PubMed] [Google Scholar]
  25. Sleytr U. B., Thornley M. J., Glauert A. M. Location of the fracture faces within the cell envelope of Acinetobacter species strain MJT-F5-5. J Bacteriol. 1974 May;118(2):693–707. doi: 10.1128/jb.118.2.693-707.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Suganuma A., Hara K., Kishida T., Nakajima K., Kawamata J. Cytological changes of Escherichia coli caused by polymyxin E. Biken J. 1968 Jun;11(2):149–155. [PubMed] [Google Scholar]
  27. Teuber M., Bader J. Quantitative correlation of uptake with antibiotic activity of polymyxin B in Salmonella typhimurium. FEBS Lett. 1971 Aug 15;16(3):195–197. doi: 10.1016/0014-5793(71)80131-x. [DOI] [PubMed] [Google Scholar]
  28. Teuber M., Bader J., Schindler P., Bowman B. F. Wirkung des Polymyxin B auf die Membranen und Membran-komponenten von Salmonella typhimurium. Zentralbl Bakteriol Orig A. 1974;228(1):243–245. [PubMed] [Google Scholar]
  29. Teuber M. Lysozyme-dependent production of spheroplast-like bodies from polymyxin B-treated Salmonella typhimurium. Arch Mikrobiol. 1970;70(2):139–146. doi: 10.1007/BF00412204. [DOI] [PubMed] [Google Scholar]
  30. Teuber M. Phosphorylation of methyl-alpha-D-glucopyranoside in polymyxin B-treated Salmonella typhimurium. J Bacteriol. 1969 Dec;100(3):1417–1419. doi: 10.1128/jb.100.3.1417-1419.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thornley M. J., Sleytr U. B. Freeze-etching of the outer membranes of Pseudomonas and Acinetobacter. Arch Microbiol. 1974;100(4):409–417. doi: 10.1007/BF00446332. [DOI] [PubMed] [Google Scholar]
  32. WARREN G. H., GRAY J., YURCHENCO J. A. Effect of polymyxin on the lysis of Neisseria catarrhalis by lysozyme. J Bacteriol. 1957 Dec;74(6):788–793. doi: 10.1128/jb.74.6.788-793.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. WEBER G. Polarization of the fluorescence of macromolecules. II. Fluorescent conjugates of ovalbumin and bovine serum albumin. Biochem J. 1952 May;51(2):155–167. doi: 10.1042/bj0510155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wahn K., Lutsch G., Rockstroh T., Zapf K. Morphological and physiological investigations on the action of polymyxin B on Escherichia coli. Arch Mikrobiol. 1968;63(2):103–116. doi: 10.1007/BF00412165. [DOI] [PubMed] [Google Scholar]
  35. van Gool A. P., Nanninga N. Fracture faces in the cell envelope of Escherichia coli. J Bacteriol. 1971 Oct;108(1):474–481. doi: 10.1128/jb.108.1.474-481.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES