Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1978 Oct;136(1):391–401. doi: 10.1128/jb.136.1.391-401.1978

Cell envelope alterations in antibiotic-sensitive and-resistant strains of Neisseria gonorrhoeae.

L F Guymon, D L Walstad, P F Sparling
PMCID: PMC218671  PMID: 101519

Abstract

The cell envelopes of antibiotic-resistant and -sensitive isogenic strains of Neisseria gonorrhoeae were analyzed to determine whether acquisition of genetic loci for altered antibiotic sensitivity was accompanied by alterations in cell envelope composition. No differences in the composition of phospholipids and lipopolysaccharides were noted. Acquisition of mtr-2, which results in low-level, nonspecific increased resistance to multiple antibiotics, dyes, and detergents, was accompanied by a sevenfold increase in the amount of a minor, 52,000-molecular-weight outer membrane protein and a 32% increase in the extent of peptidoglycan cross-linking. Subsequent addition of the nonspecific hypersensitivity loci env-1 or env-2 to a strain carrying mtr-2 resulted in reversal of the phenotypic resistance determined by mtr-2 and marked reduction in both the amount of the 52,000-molecular-weight outer membrane protein and the extent of peptidoglycan cross-linking. Introduction of penB2, which results in a fourfold increase in resistance to penicillin and tetracycline, was accompanied by the disappearance of the principal outer membrane protein of the wild-type strain (molecular weight, 36,900) and the appearance of a new species of the principal outer membrane protein (molecular weight, 39,400) in the transformant.

Full text

PDF
391

Images in this article

397Image
on p.397

Selected References

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

  1. Ames G. F., Spudich E. N., Nikaido H. Protein composition of the outer membrane of Salmonella typhimurium: effect of lipopolysaccharide mutations. J Bacteriol. 1974 Feb;117(2):406–416. doi: 10.1128/jb.117.2.406-416.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arvidson G. A. Structural and metabolic heterogeneity of rat liver glycerophosphatides. Eur J Biochem. 1968 May;4(4):478–486. doi: 10.1111/j.1432-1033.1968.tb00237.x. [DOI] [PubMed] [Google Scholar]
  3. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  4. Biswas G., Comer S., Sparling P. F. Chromosomal location of antibiotic resistance genes in Neisseria gonorrhoeae. J Bacteriol. 1976 Mar;125(3):1207–1210. doi: 10.1128/jb.125.3.1207-1210.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bragg P. D., Hou C. Organization of proteins in the native and reformed outer membrane of Escherichia coli. Biochim Biophys Acta. 1972 Aug 9;274(2):478–488. doi: 10.1016/0005-2736(72)90193-9. [DOI] [PubMed] [Google Scholar]
  6. Burman L. G., Nordström K., Bloom G. D. Murein and the outer penetration barrier of Escherichia coli K-12, Proteus mirabilis, and Pseudomonas aeruginosa. J Bacteriol. 1972 Dec;112(3):1364–1374. doi: 10.1128/jb.112.3.1364-1374.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Catlin B. W. Nutritional profiles of Neisseria gonorrhoeae, Neisseria meningitidis, and Neisseria lactamica in chemically defined media and the use of growth requirements for gonococcal typing. J Infect Dis. 1973 Aug;128(2):178–194. doi: 10.1093/infdis/128.2.178. [DOI] [PubMed] [Google Scholar]
  8. Davis R. H., Salton M. R. Some properties of a D-alanine carboxypeptidase in envelope fractions of Neisseria gonorrhoeae. Infect Immun. 1975 Nov;12(5):1065–1069. doi: 10.1128/iai.12.5.1065-1069.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dunnick J. K., O'Leary W. M. Correlation of bacteria lipid composition with antibiotic resistance. J Bacteriol. 1970 Mar;101(3):892–900. doi: 10.1128/jb.101.3.892-900.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eisenstein B. I., Sparling P. F. Mutations to increased antibiotic sensitivity in naturally-occurring gonococci. Nature. 1978 Jan 19;271(5642):242–244. doi: 10.1038/271242a0. [DOI] [PubMed] [Google Scholar]
  11. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  12. GARBUS J., DELUCA H. F., LOOMANS M. E., STRONG F. M. The rapid incorporation of phosphate into mitochondrial lipids. J Biol Chem. 1963 Jan;238:59–63. [PubMed] [Google Scholar]
  13. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  14. Guymon L. F., Sparling P. F. Altered crystal violet permeability and lytic behavior in antibiotic-resistant and -sensitive mutants of Neisseria gonorrhoeae. J Bacteriol. 1975 Nov;124(2):757–763. doi: 10.1128/jb.124.2.757-763.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hebeler B. H., Young F. E. Chemical composition and turnover of peptidoglycan in Neisseria gonorrhoeae. J Bacteriol. 1976 Jun;126(3):1180–1185. doi: 10.1128/jb.126.3.1180-1185.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hildebrandt J. F., Mayer L. W., Wang S. P., Buchanan T. M. Neisseria gonorrhoeae acquire a new principal outer-membrane protein when transformed to resistance to serum bactericidal activity. Infect Immun. 1978 Apr;20(1):267–272. doi: 10.1128/iai.20.1.267-272.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Inouye H., Beckwith J. Synthesis and processing of an Escherichia coli alkaline phosphatase precursor in vitro. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1440–1444. doi: 10.1073/pnas.74.4.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Inouye S., Wang S., Sekizawa J., Halegoua S., Inouye M. Amino acid sequence for the peptide extension on the prolipoprotein of the Escherichia coli outer membrane. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1004–1008. doi: 10.1073/pnas.74.3.1004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jarvis D., Strominger J. L. Structure of the cell wall of Staphylococcus aureus. 8. Structure and chemical synthesis of the basic peptides released by the Myxobacterium enzyme. Biochemistry. 1967 Aug;6(8):2591–2598. doi: 10.1021/bi00860a042. [DOI] [PubMed] [Google Scholar]
  20. Johnston K. H., Gotschlich E. C. Isolation and characterization of the outer membrane of Neisseria gonorrhoeae. J Bacteriol. 1974 Jul;119(1):250–257. doi: 10.1128/jb.119.1.250-257.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Johnston K. H., Holmes K. K., Gotschlich E. C. The serological classification of Neisseria gonorrhoeae. I. Isolation of the outer membrane complex responsible for serotypic specificity. J Exp Med. 1976 Apr 1;143(4):741–758. doi: 10.1084/jem.143.4.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kamiryo T., Strominger J. L. Penicillin-resistant temperature-sensitive mutants of Escherichia coli which synthesize hypo- or hyper-cross-linked peptidoglycan. J Bacteriol. 1974 Feb;117(2):568–577. doi: 10.1128/jb.117.2.568-577.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Leutgeb W., Schwarz U. Zur Biosynthese des formgebenden Elements der Bakterienzellwand. I. Abbau des Mureins als erster Schritt beim Wachstum des Sacculus. Z Naturforsch B. 1967 May;22(5):545–549. [PubMed] [Google Scholar]
  25. 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]
  26. MacGregor C. H., Schnaitman C. A. Alterations in the cytoplasmic membrane proteins of various chlorate-resistant mutants of Escherichia coli. J Bacteriol. 1971 Oct;108(1):564–570. doi: 10.1128/jb.108.1.564-570.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Maier T. W., Zubrzycki L., Coyle M. B., Chila M., Warner P. Genetic analysis of drug resistance in Neisseria gonorrhoeae: production of increased resistance by the combination of two antibiotic resistance loci. J Bacteriol. 1975 Nov;124(2):834–842. doi: 10.1128/jb.124.2.834-842.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Maness M. J., Sparling P. F. Multiple antibiotic resistance due to a single mutation in Neisseria gonorrhoeae. J Infect Dis. 1973 Sep;128(3):321–330. doi: 10.1093/infdis/128.3.321. [DOI] [PubMed] [Google Scholar]
  29. Manning P. A., Pugsley A. P., Reeves P. Defective growth functions in mutants of Escherichia coli K12 lacking a major outer membrane protein. J Mol Biol. 1977 Oct 25;116(2):285–300. doi: 10.1016/0022-2836(77)90217-0. [DOI] [PubMed] [Google Scholar]
  30. Matthieu J. M., Quarles R. H. Quantitative scanning of glycoproteins on polyacrylamide gels stained with periodic acid-Schiff reagent (PAS). Anal Biochem. 1973 Sep;55(1):313–316. doi: 10.1016/0003-2697(73)90321-7. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. 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]
  34. 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]
  35. OSBORN M. J. STUDIES ON THE GRAM-NEGATIVE CELL WALL. I. EVIDENCE FOR THE ROLE OF 2-KETO- 3-DEOXYOCTONATE IN THE LIPOPOLYSACCHARIDE OF SALMONELLA TYPHIMURIUM. Proc Natl Acad Sci U S A. 1963 Sep;50:499–506. doi: 10.1073/pnas.50.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Perry M. B., Daoust V. The lipopolysaccharides of Neisseria gonorrhoeae colony types 1 and 4. Can J Biochem. 1975 May;53(5):623–629. doi: 10.1139/o75-084. [DOI] [PubMed] [Google Scholar]
  37. Pugsley A. P., Schnaitman C. A. Outer membrane proteins of Escherichia coli. VII. Evidence that bacteriophage-directed protein 2 functions as a pore. J Bacteriol. 1978 Mar;133(3):1181–1189. doi: 10.1128/jb.133.3.1181-1189.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Rodriguez W., Saz A. K. Possible mechanism of decreased susceptibility of Neisseria gonorrhoeae to penicillin. Antimicrob Agents Chemother. 1975 Jun;7(6):788–792. doi: 10.1128/aac.7.6.788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sanderson K. E., MacAlister T., Costerton J. W., Cheng K. J. Permeability of lipopolysaccharide-deficient (rough) mutants of Salmonella typhimurium to antibiotics, lysozyme, and other agents. Can J Microbiol. 1974 Aug;20(8):1135–1145. doi: 10.1139/m74-176. [DOI] [PubMed] [Google Scholar]
  41. Sarubbi F. A., Jr, Blackman E., Sparling P. F. Genetic mapping of linked antibiotic resistance loci in Neisseria gonorrhoeae. J Bacteriol. 1974 Dec;120(3):1284–1292. doi: 10.1128/jb.120.3.1284-1292.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sarubbi F. A., Jr, Sparling P. F., Blackman E., Lewis E. Loss of low-level antibiotic resistance in Neisseria gonorrhoeae due to env mutations. J Bacteriol. 1975 Nov;124(2):750–756. doi: 10.1128/jb.124.2.750-756.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sekizawa J., Inouye S., Halegoua S., Inouye M. Precursors of major outer membrane proteins of Escherichia coli. Biochem Biophys Res Commun. 1977 Aug 8;77(3):1126–1133. doi: 10.1016/s0006-291x(77)80095-8. [DOI] [PubMed] [Google Scholar]
  44. Senff L. M., Wegener W. S., Brooks G. F., Finnerty W. R., Makula R. A. Phospholipid composition and phospholipase A activity of Neisseria gonorrhoeae. J Bacteriol. 1976 Aug;127(2):874–880. doi: 10.1128/jb.127.2.874-880.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  46. Sparling P. F., Sarubbi F. A., Jr, Blackman E. Inheritance of low-level resistance to penicillin, tetracycline, and chloramphenicol in Neisseria gonorrhoeae. J Bacteriol. 1975 Nov;124(2):740–749. doi: 10.1128/jb.124.2.740-749.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Stead A., Main J. S., Ward M. E., Watt P. J. Studies on lipopolysaccharides isolated from strains of Neisseria gonorrhoeae. J Gen Microbiol. 1975 May;88(1):123–131. doi: 10.1099/00221287-88-1-123. [DOI] [PubMed] [Google Scholar]
  48. Sud I. J., Feingold D. S. Phospholipids and fatty acids of Neisseria gonorrhoeae. J Bacteriol. 1975 Nov;124(2):713–717. doi: 10.1128/jb.124.2.713-717.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Suling W. J., O'Leary W. M. Lipids of antibiotic-resistant and -susceptible members of the Enterobacteriaceae. Can J Microbiol. 1977 Aug;23(8):1045–1051. doi: 10.1139/m77-156. [DOI] [PubMed] [Google Scholar]
  50. 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]
  51. Walstad D. L., Guymon L. F., Sparling P. F. Altered outer membrane protein in different colonial types of Neisseria gonorrhoeae. J Bacteriol. 1977 Mar;129(3):1623–1627. doi: 10.1128/jb.129.3.1623-1627.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Walstad D. L., Reitz R. C., Sparling P. F. Growth inhibition among strains of Neisseria gonorrhoeae due to production of inhibitory free fatty acids and lysophosphatidylethanolamine: absence of bacteriocins. Infect Immun. 1974 Sep;10(3):481–488. doi: 10.1128/iai.10.3.481-488.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wolf-Watz H., Elmros T., Normark S., Bloom G. D. Cell envelope of Neisseria gonorrhoeae: outer membrane and peptidoglycan composition of penicillin-sensitive and-resistant strains. Infect Immun. 1975 Jun;11(6):1332–1341. doi: 10.1128/iai.11.6.1332-1341.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wu H. C. Isolation and characterization of an Escherichia coli mutant with alteration in the outer membrane porteins of the cell envelope. Biochim Biophys Acta. 1972 Dec 1;290(1):274–289. doi: 10.1016/0005-2736(72)90070-3. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES