Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1980 Dec;30(3):773–780. doi: 10.1128/iai.30.3.773-780.1980

Resolution of Basic Gonococcal Outer Membrane Proteins by Nonequilibrium pH Gradient Electrophoresis

Robert B Jones 1,2, Patricia A Jemison 1, Wilbert J Newhall V 2, Richard A Haak 2
PMCID: PMC551382  PMID: 6785234

Abstract

Outer membrane proteins from opaque and transparent colonial variants of strain F62 of Neisseria gonorrhoeae were analyzed by two-dimensional electrophoresis with isoelectric focusing in the first dimension and sodium dodecyl sulphate-polyacrylamide gel electrophoresis in the second. Most of the higher-molecular-weight proteins focused sharply in the acidic region of the gel. In contrast, the principal outer membrane protein, a 31,000-molecular-weight protein, and the opacity-associated proteins remained near the origin (at the basic end of the gel) without focusing. However, when the samples were loaded on the acidic end of an isoelectric focusing gel and subjected to nonequilibrium pH gradient electrophoresis, these proteins behaved as basic proteins. In addition, three distinct opacity-associated heat-modifiable proteins could be identified. No other differences in the protein composition of outer membranes from opaque and transparent variants were apparent. Amino acid analysis of the principal outer membrane protein indicated that its net positive charge may result from partial amidation of its acidic residues. The unexpected observation that the major surface proteins of the gonococcus are basic may have implications for intragonococcal adhesion and for gonococcal interactions with mammalian cells.

Full text

PDF
773

Images in this article

775Fig. 1
on p.775
775Fig. 2
on p.775
776Fig. 4
on p.776
777Fig. 5
on p.777

Selected References

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

  1. Ames G. F., Nikaido K. Two-dimensional gel electrophoresis of membrane proteins. Biochemistry. 1976 Feb 10;15(3):616–623. doi: 10.1021/bi00648a026. [DOI] [PubMed] [Google Scholar]
  2. Capaldi R. A., Vanderkooi G. The low polarity of many membrane proteins. Proc Natl Acad Sci U S A. 1972 Apr;69(4):930–932. doi: 10.1073/pnas.69.4.930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Draper D. L., James J. F., Brooks G. F., Sweet R. L. Comparison of virulence markers of peritoneal and fallopian tube isolates with endocervical Neisseria gonorrhoeae isolates from women with acute salpingitis. Infect Immun. 1980 Mar;27(3):882–888. doi: 10.1128/iai.27.3.882-888.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Frasch C. E., Mocca L. F. Heat-modifiable outer membrane proteins of Neisseria meningitidis and their organization within the membrane. J Bacteriol. 1978 Dec;136(3):1127–1134. doi: 10.1128/jb.136.3.1127-1134.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hansen E. J., Wilson R. M., Baseman J. B. Two-dimensional gel electrophoretic comparison of proteins from virulent and avirulent strains of Mycoplasma pneumoniae. Infect Immun. 1979 May;24(2):468–475. doi: 10.1128/iai.24.2.468-475.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heckels J. E., Blackett B., Everson J. S., Ward M. E. The influence of surface charge on the attachment of Neisseria gonorrhoeae to human cells. J Gen Microbiol. 1976 Oct;96(2):359–364. doi: 10.1099/00221287-96-2-359. [DOI] [PubMed] [Google Scholar]
  7. Heckels J. E. The surface of Neisseria gonorrhoeae: isolation of the major components of the outer membrane. J Gen Microbiol. 1977 Apr;99(2):333–341. doi: 10.1099/00221287-99-2-333. [DOI] [PubMed] [Google Scholar]
  8. Heller K. B. Apparent molecular weights of a heat-modifiable protein from the outer membrane of Escherichia coli in gels with different acrylamide concentrations. J Bacteriol. 1978 Jun;134(3):1181–1183. doi: 10.1128/jb.134.3.1181-1183.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Heller K. B. Outer membrane of Serratia marcescens: apparent molecular weights of heat-modifiable proteins in gels with different acrylamide concentrations. J Bacteriol. 1979 Jan;137(1):670–672. doi: 10.1128/jb.137.1.670-672.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Holmes K. K., Counts G. W., Beaty H. N. Disseminated gonococcal infection. Ann Intern Med. 1971 Jun;74(6):979–993. doi: 10.7326/0003-4819-74-6-979. [DOI] [PubMed] [Google Scholar]
  12. James J. F., Swanson J. Studies on gonococcus infection. XIII. Occurrence of color/opacity colonial variants in clinical cultures. Infect Immun. 1978 Jan;19(1):332–340. doi: 10.1128/iai.19.1.332-340.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Jones R. B., Buchanan T. M. Quantitative measurement of phagocytosis of Neisseria gonorrhoeae by mouse peritoneal macrophages. Infect Immun. 1978 Jun;20(3):732–738. doi: 10.1128/iai.20.3.732-738.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kellogg D. S., Jr, Cohen I. R., Norins L. C., Schroeter A. L., Reising G. Neisseria gonorrhoeae. II. Colonial variation and pathogenicity during 35 months in vitro. J Bacteriol. 1968 Sep;96(3):596–605. doi: 10.1128/jb.96.3.596-605.1968. [DOI] [PMC free article] [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. 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]
  19. Lambden P. R., Robertson J. N., Watt P. J. Biological properties of two distinct pilus types produced by isogenic variants of Neisseria gonorrhoeae P9. J Bacteriol. 1980 Jan;141(1):393–396. doi: 10.1128/jb.141.1.393-396.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Magnusson K. E., Kihlström E., Norlander L., Norqvist A., Davies J., Normark S. Effect of colony type and pH on surface charge and hydrophobicity of Neisseria gonorrhoeae. Infect Immun. 1979 Nov;26(2):397–401. doi: 10.1128/iai.26.2.397-401.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nakamura K., Mizushima S. Effects of heating in dodecyl sulfate solution on the conformation and electrophoretic mobility of isolated major outer membrane proteins from Escherichia coli K-12. J Biochem. 1976 Dec;80(6):1411–1422. doi: 10.1093/oxfordjournals.jbchem.a131414. [DOI] [PubMed] [Google Scholar]
  22. Norlander L., Davies J., Norqvist A., Normark S. Genetic basis for colonial variation in Neisseria gonorrhoeae. J Bacteriol. 1979 Jun;138(3):762–769. doi: 10.1128/jb.138.3.762-769.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  24. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  25. Rees E., Annels E. H. Gonococcal salpingitis. Br J Vener Dis. 1969 Sep;45(3):205–215. doi: 10.1136/sti.45.3.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosenbusch J. P. Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodecyl sulfate binding. J Biol Chem. 1974 Dec 25;249(24):8019–8029. [PubMed] [Google Scholar]
  27. Salit I. E., Gotschlich E. C. Gonococcal color and opacity variants: virulence for chicken embryos. Infect Immun. 1978 Nov;22(2):359–364. doi: 10.1128/iai.22.2.359-364.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schnaitman C. A. Outer membrane proteins of Escherichia coli. I. Effect of preparative conditions on the migration of protein in polyacrylamide gels. Arch Biochem Biophys. 1973 Aug;157(2):541–552. doi: 10.1016/0003-9861(73)90673-5. [DOI] [PubMed] [Google Scholar]
  29. Swanson J. Studies on gonococcus infection. XII. Colony color and opacity varienats of gonococci. Infect Immun. 1978 Jan;19(1):320–331. doi: 10.1128/iai.19.1.320-331.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Swanson J. Studies on gonococcus infection. XIV. Cell wall protein differences among color/opacity colony variants of Neisseria gonorrhoeae. Infect Immun. 1978 Jul;21(1):292–302. doi: 10.1128/iai.21.1.292-302.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tokunaga H., Tokunaga M., Nakae T. Characterization of porins from the outer membrane of Salmonella typhimurium. 1. Chemical analysis. Eur J Biochem. 1979 Apr;95(3):433–439. doi: 10.1111/j.1432-1033.1979.tb12982.x. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. 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]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES