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. 1981 Nov;148(2):426–434. doi: 10.1128/jb.148.2.426-434.1981

Increased outer membrane resistance to ethylenediaminetetraacetate and cations in novel lipid A mutants.

M Vaara
PMCID: PMC216223  PMID: 6795177

Abstract

Polymyxin-resistant pmrA mutants of Salmonella typhimurium differed from their parents in that they were resistant to tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate-lysozyme, tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate-deoxycholate, and tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate-bacitracin. Tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate released about 50% less lipopolysaccharide from the pmrA strains than from the parental strains when the bacteria were grown in L-broth containing 2 mM Ca2+. Protamine, polylysine, octapeptin, benzalkonium chloride, cold NaCl, cold MgCl2, or cold tris(hydroxymethyl)aminomethane hydrochloride (pH 7.2) caused no leakage or markedly less leakage of periplasmic beta-lactamase from a pmrA mutant than from its parent strain. pmrA mutants were more resistant than their parent strains to protamine and polylysine but not to octapeptin or benzalkonium chloride, as measured by the ability of these agents to kill the bacteria or to sensitize them to deoxycholate-induced lysis. The pmrA strains did not differ from their parent strains in sensitivity to several antibiotics, in porin function (as measured by cephaloridine diffusion across the outer membrane), or in outer membrane-associated phospholipase A activity.

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Selected References

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

  1. Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [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. Buchanan-Davidson D. J., Seastone C. V., Stahmann M. A. ACTION OF SYNTHETIC POLYLYSINE ON THE GROWTH AND PHAGOCYTOSIS OF BACTERIA IN VITRO. J Bacteriol. 1960 Nov;80(5):590–594. doi: 10.1128/jb.80.5.590-594.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coleman W. G., Jr, Leive L. Two mutations which affect the barrier function of the Escherichia coli K-12 outer membrane. J Bacteriol. 1979 Sep;139(3):899–910. doi: 10.1128/jb.139.3.899-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Corwin L. M., Rothman S. W., Kim R., Talevi L. A. Mechanisms and genetics of resistance to sodium lauryl sulfate in strains of Shigella and Escherichia coli. Infect Immun. 1971 Sep;4(3):287–294. doi: 10.1128/iai.4.3.287-294.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Corwin L. M., Talevi L. A. Mutation in Shigella flexneri 2a resulting in increased electrophoretic mobility. Infect Immun. 1972 May;5(5):798–802. doi: 10.1128/iai.5.5.798-802.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dame J. B., Shapiro B. M. Use of polymyxin B, levallorphan, and tetracaine to isolate novel envelope mutants of Escherichia coli. J Bacteriol. 1976 Aug;127(2):961–972. doi: 10.1128/jb.127.2.961-972.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Doi O., Oki M., Nojima S. Two kinds of phospholipase A and lysophospholipase in Escherichia coli. Biochim Biophys Acta. 1972 Feb 21;260(2):244–258. [PubMed] [Google Scholar]
  9. Foulds J. TolF locus in Escherichia coli: chromosomal location and relationship to loci cmlB and tolD. J Bacteriol. 1976 Nov;128(2):604–608. doi: 10.1128/jb.128.2.604-608.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Galanos C., Lüderitz O. Electrodialysis of lipopolysaccharides and their conversion to uniform salt forms. Eur J Biochem. 1975 Jun;54(2):603–610. doi: 10.1111/j.1432-1033.1975.tb04172.x. [DOI] [PubMed] [Google Scholar]
  11. Geyer R., Galanos C., Westphal O., Golecki J. R. A lipopolysaccharide-binding cell-surface protein from Salmonella minnesota. Isolation, partial characterization and occurrence in different Enterobacteriaceae. Eur J Biochem. 1979 Jul;98(1):27–38. doi: 10.1111/j.1432-1033.1979.tb13156.x. [DOI] [PubMed] [Google Scholar]
  12. Gmeiner J., Bergmann H., Schlecht S. Molecular organization of the outer membrane of Salmonella typhimurium. Different release of lipopolysaccharide from wild type and lipopolysaccharide mutant cells by EDTA treatment. Arch Microbiol. 1980 Jan;124(1):69–71. doi: 10.1007/BF00407030. [DOI] [PubMed] [Google Scholar]
  13. Hardaway K. L., Buller C. S. Effect of ethylenediaminetetraacetate on phospholipids and outer membrane function in Escherichia coli. J Bacteriol. 1979 Jan;137(1):62–68. doi: 10.1128/jb.137.1.62-68.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hirota Y., Suzuki H., Nishimura Y., Yasuda S. On the process of cellular division in Escherichia coli: a mutant of E. coli lacking a murein-lipoprotein. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1417–1420. doi: 10.1073/pnas.74.4.1417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. KATCHALSKI E., BICHOWSKI-SLOMNITZKI L., VOLCANI B. E. The action of some water-soluble poly-alpha-amino acids on bacteria. Biochem J. 1953 Nov;55(4):671–680. doi: 10.1042/bj0550671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kusano T., Izaki K., Takahashi H. In vivo activation by polymyxin B of phospholipase from Pseudomonas aeruginosa and Escherichia coli. J Antibiot (Tokyo) 1976 Jun;29(6):674–675. doi: 10.7164/antibiotics.29.674. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lehmann V., Rupprecht E. Microheterogeneity in lipid A demonstrated by a new intermediate in the biosynthesis of 3-deozy-D-manno-octulosonic-acid--lipid A. Eur J Biochem. 1977 Dec;81(3):443–452. doi: 10.1111/j.1432-1033.1977.tb11969.x. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  23. Meyers E., Parker W. L., Brown W. E., Linnett P., Strominger J. L. EM49: a new polypeptide antibiotic active against cell membranes. Ann N Y Acad Sci. 1974 May 10;235(0):493–501. doi: 10.1111/j.1749-6632.1974.tb43286.x. [DOI] [PubMed] [Google Scholar]
  24. Morrison D. C., Jacobs D. M. Binding of polymyxin B to the lipid A portion of bacterial lipopolysaccharides. Immunochemistry. 1976 Oct;13(10):813–818. doi: 10.1016/0019-2791(76)90181-6. [DOI] [PubMed] [Google Scholar]
  25. Mühlradt P. F., Wray V., Lehmann V. A 31P-nuclear-magnetic-resonance study of the phosphate groups in lipopolysaccharide and lipid A from Salmonella. Eur J Biochem. 1977 Nov 15;81(1):193–203. doi: 10.1111/j.1432-1033.1977.tb11941.x. [DOI] [PubMed] [Google Scholar]
  26. NETER E., GORZYNSKI E. A., WESTPHAL O., LUDERITZ O., KLUMPP D. J. The effects of protamine and histone on enterobacterial lipopolysaccharides and hemolysis. Can J Microbiol. 1958 Aug;4(4):371–383. doi: 10.1139/m58-039. [DOI] [PubMed] [Google Scholar]
  27. Nakamura H. Gene-Controlled Resistance to Acriflavine and Other Basic Dyes in Escherichia coli. J Bacteriol. 1965 Jul;90(1):8–14. doi: 10.1128/jb.90.1.8-14.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nikaido H., Nakae T. The outer membrane of Gram-negative bacteria. Adv Microb Physiol. 1979;20:163–250. doi: 10.1016/s0065-2911(08)60208-8. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Nolan J. P., Ali M. V. Effect of cholestyramine on endotoxin toxicity and absorption. Am J Dig Dis. 1972 Feb;17(2):161–166. doi: 10.1007/BF02232738. [DOI] [PubMed] [Google Scholar]
  31. Normark S., Boman H. G., Matsson E. Mutant of Escherichia coli with anomalous cell division and ability to decrease episomally and chromosomally mediated resistance to ampicillin and several other antibiotics. J Bacteriol. 1969 Mar;97(3):1334–1342. doi: 10.1128/jb.97.3.1334-1342.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Roantree R. J., Kuo T. T., MacPhee D. G. The effect of defined lipopolysaccharide core defects upon antibiotic resistances of Salmonella typhimurium. J Gen Microbiol. 1977 Dec;103(2):223–234. doi: 10.1099/00221287-103-2-223. [DOI] [PubMed] [Google Scholar]
  34. SKARNES R. C., WATSON D. W. Antimicrobial factors of normal tissues and fluids. Bacteriol Rev. 1957 Dec;21(4):273–294. doi: 10.1128/br.21.4.273-294.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sargent M. G. Rapid fixed-time assay for penicillinase. J Bacteriol. 1968 Apr;95(4):1493–1494. doi: 10.1128/jb.95.4.1493-1494.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schindler M., Osborn M. J. Interaction of divalent cations and polymyxin B with lipopolysaccharide. Biochemistry. 1979 Oct 2;18(20):4425–4430. doi: 10.1021/bi00587a024. [DOI] [PubMed] [Google Scholar]
  37. Schlecht S., Schmidt G. Möglichkeiten zur Differenzierung von Salmonella-R-Formen mittels Antibiotica und antibakterieller Farbstoffe. Zentralbl Bakteriol Orig. 1970;212(2):505–511. [PubMed] [Google Scholar]
  38. Storm D. R., Rosenthal K. S., Swanson P. E. Polymyxin and related peptide antibiotics. Annu Rev Biochem. 1977;46:723–763. doi: 10.1146/annurev.bi.46.070177.003451. [DOI] [PubMed] [Google Scholar]
  39. Sud I. J., Feingold D. S. Effect of polymyxin B on antibiotic-resistant Proteus mirabilis. Antimicrob Agents Chemother. 1972 May;1(5):417–421. doi: 10.1128/aac.1.5.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Vaara M., Vaara T. Outer membrane permeability barrier disruption by polymyxin in polymyxin-susceptible and -resistant Salmonella typhimurium. Antimicrob Agents Chemother. 1981 Apr;19(4):578–583. doi: 10.1128/aac.19.4.578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Vaara M., Vaara T., Sarvas M. Decreased binding of polymyxin by polymyxin-resistant mutants of Salmonella typhimurium. J Bacteriol. 1979 Aug;139(2):664–667. doi: 10.1128/jb.139.2.664-667.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Van Alphen L., Lugtenberg B., Rietschel E. T., Mombers C. Architecture of the outer membrane of Escherichia coli K12. Phase transitions of the bacteriophage K3 receptor complex. Eur J Biochem. 1979 Nov;101(2):571–579. doi: 10.1111/j.1432-1033.1979.tb19752.x. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. WEISSBACH A., HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed] [Google Scholar]
  45. Wilkinson R. G., Gemski P., Jr, Stocker B. A. Non-smooth mutants of Salmonella typhimurium: differentiation by phage sensitivity and genetic mapping. J Gen Microbiol. 1972 May;70(3):527–554. doi: 10.1099/00221287-70-3-527. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. van Alphen L., Verkleij A., Leunissen-Bijvelt J., Lugtenberg B. Architecture of the outer membrane of Escherichia coli. III. Protein-lipopolysaccharide complexes in intramembraneous particles. J Bacteriol. 1978 Jun;134(3):1089–1098. doi: 10.1128/jb.134.3.1089-1098.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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