Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1981 Jul;33(1):212–222. doi: 10.1128/iai.33.1.212-222.1981

Effects of proteolytic enzymes on the outer membrane proteins of Neisseria gonorrhoeae.

M S Blake, E C Gotschlich, J Swanson
PMCID: PMC350679  PMID: 6790441

Abstract

Proteolytic enzymes inhibit the growth of some strains and opacity variants of Neisseria gonorrhoeae. To understand the inhibitory effects of these enzymes, we examined several strains to determine the actions of proteases on the three predominant proteins in gonococcal outer membranes. namely, the major outer membrane protein (protein I), the sometimes-expressed opaque protein (protein II), and protein III. In a comparison of the protein I species expressed by different strains, we observed a pattern based on subunit molecular weight and susceptibility to enzymatic degradation. Protein I species having molecular weights of 34,000 were more susceptible to proteolysis, whereas protein I species having molecular weights of 33,000 were less susceptible, and protein I species having molecular weights of 32,000 were resistant. This pattern was observed both in intact cells and in purified outer membranes. All of the enzymes degraded protein II, but this susceptibility appeared to be influenced in part by the species of protein I present. Protein III was resistant to all of the proteolytic enzymes tested. Based on the resulting fragments from each proteolytic cleavage of proteins I and II and their membrane associations, we suggest how these proteins may be arranged in intact membranes. Our data suggested the presence of an endogenous gonococcal enzyme. This enzyme appeared to degrade proteins I and II into fragments resembling the fragments resulting from the action of chymotrypsin.

Full text

PDF
212

Images in this article

215Image
on p.215
216Image
on p.216
217Image
on p.217
218Image
on p.218
219Image
on p.219
220Image
on p.220

Selected References

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

  1. CASTANEDA-AGULLO M., DEL CASTILLO L. M. Simultaneous hydrolyses of esters and proteins at saturation levels. J Gen Physiol. 1958 Jan 20;41(3):485–500. doi: 10.1085/jgp.41.3.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CASTANEDA-AGULLO M., DEL CASTILLO L. M. The action of alpha chymotrypsin upon mixtures of esters and proteins at enzyme-saturating concentrations. J Gen Physiol. 1958 Sep 20;42(1):49–68. doi: 10.1085/jgp.42.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cannon J. G., Klapper D. G., Blackman E. Y., Sparling P. F. Genetic locus (nmp-1) affecting the principal outer membrane protein of Neisseria gonorrhoeae. J Bacteriol. 1980 Aug;143(2):847–851. doi: 10.1128/jb.143.2.847-851.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. 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]
  7. 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]
  8. 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]
  9. 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]
  10. KELLOGG D. S., Jr, PEACOCK W. L., Jr, DEACON W. E., BROWN L., PIRKLE D. I. NEISSERIA GONORRHOEAE. I. VIRULENCE GENETICALLY LINKED TO CLONAL VARIATION. J Bacteriol. 1963 Jun;85:1274–1279. doi: 10.1128/jb.85.6.1274-1279.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Lambden P. R., Heckels J. E., James L. T., Watt P. J. Variations in surface protein composition associated with virulence properties in opacity types of Neisseria gonorrhoeae. J Gen Microbiol. 1979 Oct;114(2):305–312. doi: 10.1099/00221287-114-2-305. [DOI] [PubMed] [Google Scholar]
  14. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  15. McDade R. L., Jr, Johnston K. H. Characterization of serologically dominant outer membrane proteins of Neisseria gonorrhoeae. J Bacteriol. 1980 Mar;141(3):1183–1191. doi: 10.1128/jb.141.3.1183-1191.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Spector T. Refinement of the coomassie blue method of protein quantitation. A simple and linear spectrophotometric assay for less than or equal to 0.5 to 50 microgram of protein. Anal Biochem. 1978 May;86(1):142–146. doi: 10.1016/0003-2697(78)90327-5. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Wong T. P., Shockley R. K., Johnston K. H. WSJM, a simple chemically defined medium for growth of Neisseria gonorrhoeae. J Clin Microbiol. 1980 Apr;11(4):363–369. doi: 10.1128/jcm.11.4.363-369.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES