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. 1980 Jan;141(1):169–176. doi: 10.1128/jb.141.1.169-176.1980

Chemical Analysis of Major Outer Membrane Proteins of Neisseria meningitidis: Comparison of Serotypes 2 and 11

Chao-Ming Tsai 1, Carl E Frasch 1
PMCID: PMC293555  PMID: 6766438

Abstract

Most of the 15 protein serotypes found in group B Neisseria meningitidis have distinct major outer membrane protein patterns when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) by the Weber-Osborn system. Both serotypes 2 and 11 contain major outer membrane proteins with apparent molecular weights of 41,000 and 28,000 (41K and 28K). The 41K and 28K proteins were purified from the prototype strains of these two serotypes (M986 type 2 and M136 type 11) by preparative slab SDS-PAGE and were chemically characterized. No hexosamine was found in the purified 41K and 28K proteins. Although the two 41K proteins had similar amino acid compositions, their mobilities in Laemmli SDS-PAGE and their fragmentation patterns on SDS-PAGE after cyanogen bromide cleavage were different. The two 28K proteins differed in their amino acid composition, mobilities in Laemmli SDS-PAGE, and cyanogen bromide cleavage products. Peptide maps following chymotrypsin digestion of radioiodinated 41K and 28K proteins revealed distinct peptide maps for all four proteins. Comparison of the peptide maps of two 41K or two 28K proteins indicated that most of the unique peptides were hydrophilic, whereas most of the common peptides were hydrophobic. These results indicated that both of the 41K proteins and the 28K proteins from serotypes 2 and 11 were chemically different, although the proteins having the same molecular weights appeared to share common peptides.

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Selected References

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

  1. BRANHAM S. E. Serological relationships among meningococci. Bacteriol Rev. 1953 Sep;17(3):175–188. doi: 10.1128/br.17.3.175-188.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bassford P. J., Jr, Diedrich D. L., Schnaitman C. L., Reeves P. Outer membrane proteins of Escherichia coli. VI. Protein alteration in bacteriophage-resistant mutants. J Bacteriol. 1977 Aug;131(2):608–622. doi: 10.1128/jb.131.2.608-622.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Craven D. E., Frasch C. E., Robbins J. B., Feldman H. A. Serogroup identification of Neisseria meningitidis: comparison of an antiserum agar method with bacterial slide agglutination. J Clin Microbiol. 1978 May;7(5):410–414. doi: 10.1128/jcm.7.5.410-414.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Diedrich D. L., Summers A. O., Schnaitman C. A. Outer membrane proteins of Escherichia coli. V. Evidence that protein 1 and bacteriophage-directed protein 2 are different polypeptides. J Bacteriol. 1977 Aug;131(2):598–607. doi: 10.1128/jb.131.2.598-607.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Elder J. H., Pickett R. A., 2nd, Hampton J., Lerner R. A. Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein. J Biol Chem. 1977 Sep 25;252(18):6510–6515. [PubMed] [Google Scholar]
  7. 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]
  8. Frasch C. E., Chapman S. S. Classification of Neisseria meningitidis group B into distinct serotypes. IV. Preliminary chemical studies on the nature of the serotype antigen. Infect Immun. 1972 Nov;6(5):674–681. doi: 10.1128/iai.6.5.674-681.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Frasch C. E., Gotschlich E. C. An outer membrane protein of Neisseria meningitidis group B responsible for serotype specificity. J Exp Med. 1974 Jul 1;140(1):87–104. doi: 10.1084/jem.140.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frasch C. E., McNelis R. M., Gotschlich E. C. Strain-specific variation in the protein and lipopolysaccharide composition of the group B meningococcal outer membrane. J Bacteriol. 1976 Aug;127(2):973–981. doi: 10.1128/jb.127.2.973-981.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GROSS E., WITKOP B. Nonenzymatic cleavage of peptide bonds: the methionine residues in bovine pancreatic ribonuclease. J Biol Chem. 1962 Jun;237:1856–1860. [PubMed] [Google Scholar]
  12. 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]
  13. Jennings H. J., Hawes G. B., Adams G. A., Kenny C. P. The chemical composition and serological reactions of lipopolysaccharides from serogroups A,B,X, and Y Neisseria meningitidis. Can J Biochem. 1973 Oct;51(10):1347–1354. doi: 10.1139/o73-178. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Mandrell R. E., Zollinger W. D. Lipopolysaccharide serotyping of Neisseria meningitidis by hemagglutination inhibition. Infect Immun. 1977 May;16(2):471–475. doi: 10.1128/iai.16.2.471-475.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nielsen T. B., Reynolds J. A. Measurements of molecular weights by gel electrophoresis. Methods Enzymol. 1978;48:3–10. doi: 10.1016/s0076-6879(78)48003-6. [DOI] [PubMed] [Google Scholar]
  18. Ozols J., Gerard C. Cleavage of tryptophanyl peptide bonds in cytochrome b5 by cyanogen bromide. J Biol Chem. 1977 Sep 10;252(17):5986–5989. [PubMed] [Google Scholar]
  19. Reynolds J. A., Tanford C. Binding of dodecyl sulfate to proteins at high binding ratios. Possible implications for the state of proteins in biological membranes. Proc Natl Acad Sci U S A. 1970 Jul;66(3):1002–1007. doi: 10.1073/pnas.66.3.1002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schroeder W. A., Shelton J. B., Shelton J. R. An examination of conditions for the cleavage of polypeptide chains with cyanogen bromide: application to catalase. Arch Biochem Biophys. 1969 Mar;130(1):551–556. doi: 10.1016/0003-9861(69)90069-1. [DOI] [PubMed] [Google Scholar]
  21. Simpson R. J., Neuberger M. R., Liu T. Y. Complete amino acid analysis of proteins from a single hydrolysate. J Biol Chem. 1976 Apr 10;251(7):1936–1940. [PubMed] [Google Scholar]
  22. Swank R. T., Munkres K. D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971 Feb;39(2):462–477. doi: 10.1016/0003-2697(71)90436-2. [DOI] [PubMed] [Google Scholar]
  23. Swanson J. Studies on gonococcus infection. XVIII. 125I-labeled peptide mapping of the major protein of the gonococcal cell wall outer membrane. Infect Immun. 1979 Mar;23(3):799–810. doi: 10.1128/iai.23.3.799-810.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]

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