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. 1972 Nov;6(5):835–851. doi: 10.1128/iai.6.5.835-851.1972

Isolation and Characterization of a Native Cell Wall Complex from Neisseria meningitidis

W D Zollinger 1, D L Kasper 1, B J Veltri 1, M S Artenstein 1
PMCID: PMC422619  PMID: 4629210

Abstract

A cell wall complex has been isolated by gentle methods from both the medium supernatant fluid and whole organisms of Neissieria meningitidis cultures. The two types of preparations have been shown to be essentially identical on the basis of chemical composition, electron microscopy, and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Four major components were identified in the complex: group-specific polysaccharide (4 to 10%), protein (45 to 65%), lipopolysaccharide (10 to 25%), and lipid (15 to 30%). The whole complex was found to be immunogenic in rabbits and to elicit production of antibody directed against the protein, the group-specific polysaccharide, and the lipopolysaccharide components. The isolated protein component was also found to be immunogenic in rabbits and to elicit production of serotype-specific antibody. The protein component was found to produce a band pattern in SDS-PAGE that is simple, reproducible, and strain dependent. The lipopolysaccharide component was found to have chemical and biological properties characteristic of bacterial endotoxin. We propose that this complex is representative of the outer trilaminar membrane of the meningococcal cell envelope in its native state.

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Selected References

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  1. Artenstein M. S., Brandt B. L., Tramont E. C., Branche W. C., Jr, Fleet H. D., Cohen R. L. Serologic studies of meningococcal infection and polysaccharide vaccination. J Infect Dis. 1971 Sep;124(3):277–288. doi: 10.1093/infdis/124.3.277. [DOI] [PubMed] [Google Scholar]
  2. Counts G. W., Seeley L., Beaty H. N. Identification of an epidemic strain of Neisseria meningitidis by bacteriocin typing. J Infect Dis. 1971 Jul;124(1):26–32. doi: 10.1093/infdis/124.1.26. [DOI] [PubMed] [Google Scholar]
  3. Frasch C. E., Chapman S. S. Classification of Neisseria meningitidis group B into distinct serotypes. I. Serological typing by a microbactericidal method. Infect Immun. 1972 Jan;5(1):98–102. doi: 10.1128/iai.5.1.98-102.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gold R., Wyle F. A. New Classification of Neisseria meningitidis by Means of Bactericidal Reactions. Infect Immun. 1970 May;1(5):479–484. doi: 10.1128/iai.1.5.479-484.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gotschlich E. C., Liu T. Y., Artenstein M. S. Human immunity to the meningococcus. 3. Preparation and immunochemical properties of the group A, group B, and group C meningococcal polysaccharides. J Exp Med. 1969 Jun 1;129(6):1349–1365. doi: 10.1084/jem.129.6.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ikawa M. Studies on a lipopolysaccharide from Escherichia coli. Ann N Y Acad Sci. 1966 Jun 30;133(2):476–485. doi: 10.1111/j.1749-6632.1966.tb52384.x. [DOI] [PubMed] [Google Scholar]
  7. Kasper D. L., Wyle F. A., Formal S. B. Bactericidal antibody assay using 14 C-labeled Neisseria meningitidis. Proc Soc Exp Biol Med. 1972 Apr;139(4):1175–1180. doi: 10.3181/00379727-139-36324. [DOI] [PubMed] [Google Scholar]
  8. Kingsbury D. T. Bacteriocin production by strains of Neisseria meningitidis. J Bacteriol. 1966 May;91(5):1696–1699. doi: 10.1128/jb.91.5.1696-1699.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Knox K. W., Cullen J., Work E. An extracellular lipopolysaccharide-phospholipid-protein complex produced by Escherichia coli grown under lysine-limiting conditions. Biochem J. 1967 Apr;103(1):192–201. doi: 10.1042/bj1030192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Leive L. Release of lipopolysaccharide by EDTA treatment of E. coli. Biochem Biophys Res Commun. 1965 Nov 22;21(4):290–296. doi: 10.1016/0006-291x(65)90191-9. [DOI] [PubMed] [Google Scholar]
  12. Liu T. Y., Gotschlich E. C., Dunne F. T., Jonssen E. K. Studies on the meningococcal polysaccharides. II. Composition and chemical properties of the group B and group C polysaccharide. J Biol Chem. 1971 Aug 10;246(15):4703–4712. [PubMed] [Google Scholar]
  13. Liu T. Y., Gotschlich E. C., Jonssen E. K., Wysocki J. R. Studies on the meningococcal polysaccharides. I. Composition and chemical properties of the group A polysaccharide. J Biol Chem. 1971 May 10;246(9):2849–2858. [PubMed] [Google Scholar]
  14. MERGENHAGEN S. E., MARTIN G. R., SCHIFFMANN E. Studies on an endotoxin of a group C Neisseria meningitidis. J Immunol. 1963 Feb;90:312–317. [PubMed] [Google Scholar]
  15. Mergenhagen S. E., Bladen H. A., Hsu K. C. Electron microscopic localization of endotoxic lipopolysaccharide in gram-negative organisms. Ann N Y Acad Sci. 1966 Jun 30;133(2):279–291. doi: 10.1111/j.1749-6632.1966.tb52371.x. [DOI] [PubMed] [Google Scholar]
  16. Neville D. M., Jr, Glossmann H. Plasma membrane protein subunit composition. A comparative study by discontinuous electrophoresis in sodium dodecyl sulfate. J Biol Chem. 1971 Oct 25;246(20):6335–6338. [PubMed] [Google Scholar]
  17. 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]
  18. Okuda S., Weinbaum G. An envelope-specific glycoprotein from Escherichia coli B. Biochemistry. 1968 Aug;7(8):2819–2825. doi: 10.1021/bi00848a018. [DOI] [PubMed] [Google Scholar]
  19. RIBI E., HASKINS W. T., LANDY M., MILNER K. C. Preparation and host-reactive properties of endotoxin with low content of nitrogen and lipid. J Exp Med. 1961 Nov 1;114:647–663. doi: 10.1084/jem.114.5.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. ROE J. H. The determination of sugar in blood and spinal fluid with anthrone reagent. J Biol Chem. 1955 Jan;212(1):335–343. [PubMed] [Google Scholar]
  21. RYTER A., KELLENBERGER E., BIRCHANDERSEN A., MAALOE O. Etude au microscope électronique de plasmas contenant de l'acide désoxyribonucliéique. I. Les nucléoides des bactéries en croissance active. Z Naturforsch B. 1958 Sep;13B(9):597–605. [PubMed] [Google Scholar]
  22. Roberts R. B. The interaction in vitro between group B meningococci and rabbit polymorphonuclear leukocytes. Demonstration of type specific opsonins and bactericidins. J Exp Med. 1967 Nov 1;126(5):795–818. doi: 10.1084/jem.126.5.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rogers S. W., Gilleland H. E., Jr, Eagon R. G. Characterization of a protein-lipopolysaccharide complex released from cell walls of Pseudomonas aeruginosa by ethylenediaminetetraacetic acid. Can J Microbiol. 1969 Jul;15(7):743–748. doi: 10.1139/m69-130. [DOI] [PubMed] [Google Scholar]
  24. Rothfield L., Pearlman-Kothencz M. Synthesis and assembly of bacterial membrane components. A lipopolysaccharide-phospholipid-protein complex excreted by living bacteria. J Mol Biol. 1969 Sep 28;44(3):477–492. doi: 10.1016/0022-2836(69)90374-x. [DOI] [PubMed] [Google Scholar]
  25. SMITH R. T., THOMAS L. The lethal effect of endotoxins on the chick embryo. J Exp Med. 1956 Aug 1;104(2):217–231. doi: 10.1084/jem.104.2.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SNYDER F., STEPHENS N. A simplified spectrophotometric determination of ester groups in lipids. Biochim Biophys Acta. 1959 Jul;34:244–245. doi: 10.1016/0006-3002(59)90255-0. [DOI] [PubMed] [Google Scholar]
  27. SVENNERHOLM L. Quantitative estimation of sialic acids. II. A colorimetric resorcinol-hydrochloric acid method. Biochim Biophys Acta. 1957 Jun;24(3):604–611. doi: 10.1016/0006-3002(57)90254-8. [DOI] [PubMed] [Google Scholar]
  28. Shands J. W. Localization of somatic antigen on gram-negative bacteria using ferritin antibody conjugates. Ann N Y Acad Sci. 1966 Jun 30;133(2):292–298. doi: 10.1111/j.1749-6632.1966.tb52372.x. [DOI] [PubMed] [Google Scholar]
  29. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  30. WATSON R. G., SCHERP H. W. The specific hapten of group C (group II alpha) meningococcus. I. Preparation and immunological behavior. J Immunol. 1958 Oct;81(4):331–336. [PubMed] [Google Scholar]
  31. 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]
  32. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  33. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

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