Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1979 Feb;15(2):182–189. doi: 10.1128/aac.15.2.182

Outer-Membrane Penetration Barriers as Components of Intrinsic Resistance to Beta-Lactam and Other Antibiotics in Escherichia coli K-12

R Allan Scudamore 1, Terrance J Beveridge 2, Morris Goldner 1
PMCID: PMC352630  PMID: 106773

Abstract

A new technique has been devised to investigate the penetration of antibiotics through the gram-negative outer membrane; the application here was to study intrinsic resistance of Escherichia coli K-12. Exponential cells in broth were briefly treated with 2.5 mM ethylenediaminetetraacetic acid at 5°C to disrupt the outer membrane penetration barrier, and the response of treated and untreated cells to antibiotics was compared by turbidimetry. A barrier index was derived to describe the ability of 7 beta-lactam and 10 other antibiotics to penetrate the outer membrane of strain Y10. There was correlation between the molecular weight and log10 barrier index (r = 0.59, P ≅ 0.01). The envelope mutant D22 (envA) had low barrier indexes for erythromycin, rifampin, ampicillin, and cloxacillin. For the beta-lactams, outer membrane penetration and affinity for inner membrane target site(s) triggering cell lysis were measured as independent components of the overall activity; although penetration and overall activity varied greatly, the affinities of most were within a narrow range.

Full text

PDF
182

Selected References

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

  1. Bayer M. E., Leive L. Effect of ethylenediaminetetraacetate upon the surface of Escherichia coli. J Bacteriol. 1977 Jun;130(3):1364–1381. doi: 10.1128/jb.130.3.1364-1381.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Blumberg P. M. Penicillin binding components of bacterial cells and their relationship to the mechanism of penicillin action. Ann N Y Acad Sci. 1974 May 10;235(0):310–325. doi: 10.1111/j.1749-6632.1974.tb43274.x. [DOI] [PubMed] [Google Scholar]
  4. Boman H. G., Jonsson S., Monner D., Normark S., Bloom G. D. Cell-surface alterations in Escherichia coli K-12 with chromosmal mutations changing ampicillin resistance. Ann N Y Acad Sci. 1971 Jun 11;182:342–357. doi: 10.1111/j.1749-6632.1971.tb30670.x. [DOI] [PubMed] [Google Scholar]
  5. Boman H. G., Nordström K., Normark S. Penicillin resistance in Escherichia coli K12: synergism between penicillinases and a barrier in the outer part of the envelope. Ann N Y Acad Sci. 1974 May 10;235(0):569–586. doi: 10.1111/j.1749-6632.1974.tb43291.x. [DOI] [PubMed] [Google Scholar]
  6. Braun V., Gnirke H., Henning U., Rehn K. Model for the structure of the shape-maintaining layer of the Escherichia coli cell envelope. J Bacteriol. 1973 Jun;114(3):1264–1270. doi: 10.1128/jb.114.3.1264-1270.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Decad G. M., Nikaido H. Outer membrane of gram-negative bacteria. XII. Molecular-sieving function of cell wall. J Bacteriol. 1976 Oct;128(1):325–336. doi: 10.1128/jb.128.1.325-336.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Formanek H. A three dimensional model of the digestion of peptidoglycan by lysozyme. Biophys Struct Mech. 1977 Dec 27;4(1):1–14. doi: 10.1007/BF00538836. [DOI] [PubMed] [Google Scholar]
  9. Hamilton-Miller J. M. Effect of EDTA upon bacterial permeability to benzylpenicillin. Biochem Biophys Res Commun. 1965 Sep 22;20(6):688–691. doi: 10.1016/0006-291x(65)90070-7. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Leive L. Studies on the permeability change produced in coliform bacteria by ethylenediaminetetraacetate. J Biol Chem. 1968 May 10;243(9):2373–2380. [PubMed] [Google Scholar]
  12. 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]
  13. Lutkenhaus J. F. Role of a major outer membrane protein in Escherichia coli. J Bacteriol. 1977 Aug;131(2):631–637. doi: 10.1128/jb.131.2.631-637.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Matsushita K., Adachi O., Shinagawa E., Ameyama M. Isolation and characterization of outer and inner membranes from Pseudomonas aeruginosa and effect of EDTA on the membranes. J Biochem. 1978 Jan;83(1):171–181. doi: 10.1093/oxfordjournals.jbchem.a131888. [DOI] [PubMed] [Google Scholar]
  15. Medeiros A. A., Kent R. L., O'Brien T. F. Characterization and prevalence of the different mechanisms of resistance to beta-lactam antibiotics in clinical isolates of Escherichia coli. Antimicrob Agents Chemother. 1974 Dec;6(6):791–801. doi: 10.1128/aac.6.6.791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nakae T. Identification of the outer membrane protein of E. coli that produces transmembrane channels in reconstituted vesicle membranes. Biochem Biophys Res Commun. 1976 Aug 9;71(3):877–884. doi: 10.1016/0006-291x(76)90913-x. [DOI] [PubMed] [Google Scholar]
  17. Nakae T. Outer membrane of Salmonella typhimurium: reconstitution of sucrose-permeable membrane vesicles. Biochem Biophys Res Commun. 1975 Jun 16;64(4):1224–1230. doi: 10.1016/0006-291x(75)90823-2. [DOI] [PubMed] [Google Scholar]
  18. Nakae T. Outer membrane of Salmonella. Isolation of protein complex that produces transmembrane channels. J Biol Chem. 1976 Apr 10;251(7):2176–2178. [PubMed] [Google Scholar]
  19. 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]
  20. Nikaido H. Outer membrane of Salmonella typhimurium. Transmembrane diffusion of some hydrophobic substances. Biochim Biophys Acta. 1976 Apr 16;433(1):118–132. doi: 10.1016/0005-2736(76)90182-6. [DOI] [PubMed] [Google Scholar]
  21. Nikaido H., Song S. A., Shaltiel L., Nurminen M. Outer membrane of Salmonella XIV. Reduced transmembrane diffusion rates in porin-deficient mutants. Biochem Biophys Res Commun. 1976 May 23;76(2):324–330. doi: 10.1016/0006-291x(77)90728-8. [DOI] [PubMed] [Google Scholar]
  22. Normark S., Westling B. Nature of the penetration barrier in Escherichia coli K-12: effect of macromolecular inhibition of penetrability in strains containing the envA gene. J Bacteriol. 1971 Oct;108(1):45–50. doi: 10.1128/jb.108.1.45-50.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Park J. T., Edwards J. R., Wise E. M., Jr In vivo studies on the uptake and binding of beta-lactam antibiotics in relation to inhibition of wall synthesis and cell death. Ann N Y Acad Sci. 1974 May 10;235(0):300–309. doi: 10.1111/j.1749-6632.1974.tb43273.x. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Retsema J. A., Ray V. A. Correlation between the binding of beta-lactam antibiotics to Staphylococcus aureus and their physical-chemical properties. Antimicrob Agents Chemother. 1972 Sep;2(3):173–180. doi: 10.1128/aac.2.3.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Richmond M. H., Clark D. C., Wotton S. Indirect method for assessing the penetration of beta-lactamase-nonsusceptible penicillins and cephalosporins in Escherichia coli strains. Antimicrob Agents Chemother. 1976 Aug;10(2):215–218. doi: 10.1128/aac.10.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Salton M. R., Owen P. Bacterial membrane structure. Annu Rev Microbiol. 1976;30:451–482. doi: 10.1146/annurev.mi.30.100176.002315. [DOI] [PubMed] [Google Scholar]
  28. Spratt B. G. Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2999–3003. doi: 10.1073/pnas.72.8.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Spratt B. G. Properties of the penicillin-binding proteins of Escherichia coli K12,. Eur J Biochem. 1977 Jan;72(2):341–352. doi: 10.1111/j.1432-1033.1977.tb11258.x. [DOI] [PubMed] [Google Scholar]
  30. TREFFERS H. P. The linear representation of dosage-response curves in microbial-antibiotic assays. J Bacteriol. 1956 Jul;72(1):108–114. doi: 10.1128/jb.72.1.108-114.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tomasz A., Waks S. Mechanism of action of penicillin: triggering of the pneumococcal autolytic enzyme by inhibitors of cell wall synthesis. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4162–4166. doi: 10.1073/pnas.72.10.4162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Voll M. J., Leive L. Release of lipopolysaccharide in Escherichia coli resistant to the permeability increase induced by ethylenediaminetetraacetate. J Biol Chem. 1970 Mar 10;245(5):1108–1114. [PubMed] [Google Scholar]
  33. Zimmermann W., Rosselet A. Function of the outer membrane of Escherichia coli as a permeability barrier to beta-lactam antibiotics. Antimicrob Agents Chemother. 1977 Sep;12(3):368–372. doi: 10.1128/aac.12.3.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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