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. 1988 Apr;170(4):1866–1873. doi: 10.1128/jb.170.4.1866-1873.1988

Purification, characterization, and comparison of the immunoglobulin A1 proteases of Neisseria gonorrhoeae.

D A Simpson 1, R P Hausinger 1, M H Mulks 1
PMCID: PMC211043  PMID: 3127382

Abstract

Each isolate of Neisseria gonorrhoeae produces one of two distinct immunoglobulin A1 (IgA1) proteases, type 1 or type 2, which are known to possess different cleavage specificities for peptide bonds in the hinge region of human IgA1. Both proteases were secreted into the culture medium throughout exponential growth; however, the activity level of the type 2 protease was 10-fold that observed for the type 1 enzyme. The type 2 protease was quite stable and resistant to a variety of inhibitors. In contrast, the type 1 enzyme was highly unstable and inhibited by low concentrations of metal chelators, salts, and thiol- or serine-specific chemical reagents. Both types of gonococcal IgA1 protease were purified from broth culture supernatants by a combination of anion-exchange, chromatofocusing, and molecular sieve chromatography techniques. The stable type 2 enzyme comprised a 114-kilodalton (kDa) peptide which converted to a still active 109-kDa peptide during isolation. In contrast, the type 1 protease possessed a 112-kDa peptide which did not convert to a smaller form and which could not be dissociated from peptides of 34 and 31 kDa without complete loss of enzyme activity.

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Selected References

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

  1. Blake M. S., Swanson J. Studies on gonococcus infection. XVI. Purification of Neisseria gonorrhoeae immunoglobulin A1 protease. Infect Immun. 1978 Nov;22(2):350–358. doi: 10.1128/iai.22.2.350-358.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Bricker J., Mulks M. H., Plaut A. G., Moxon E. R., Wright A. IgA1 proteases of Haemophilus influenzae: cloning and characterization in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1983 May;80(9):2681–2685. doi: 10.1073/pnas.80.9.2681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  5. Halter R., Pohlner J., Meyer T. F. IgA protease of Neisseria gonorrhoeae: isolation and characterization of the gene and its extracellular product. EMBO J. 1984 Jul;3(7):1595–1601. doi: 10.1002/j.1460-2075.1984.tb02016.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jack G. W., Richmond M. H. A comparative study of eight distinct beta-lactamases synthesized by gram-negative bacteria. J Gen Microbiol. 1970 Apr;61(1):43–61. doi: 10.1099/00221287-61-1-43. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Kilian M., Thomsen B., Petersen T. E., Bleeg H. S. Occurrence and nature of bacterial IgA proteases. Ann N Y Acad Sci. 1983 Jun 30;409:612–624. doi: 10.1111/j.1749-6632.1983.tb26903.x. [DOI] [PubMed] [Google Scholar]
  9. Knapp J. S., Holmes K. K. Disseminated gonococcal infections caused by Neisseria gonorrhoeae with unique nutritional requirements. J Infect Dis. 1975 Aug;132(2):204–208. doi: 10.1093/infdis/132.2.204. [DOI] [PubMed] [Google Scholar]
  10. Koomey J. M., Gill R. E., Falkow S. Genetic and biochemical analysis of gonococcal IgA1 protease: cloning in Escherichia coli and construction of mutants of gonococci that fail to produce the activity. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7881–7885. doi: 10.1073/pnas.79.24.7881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Morse S. A., Bartenstein L. Purine metabolism in Neisseria gonorrhoeae: the requirement for hypoxanthine. Can J Microbiol. 1980 Jan;26(1):13–20. doi: 10.1139/m80-003. [DOI] [PubMed] [Google Scholar]
  13. Morse S. A., Stein S., Hines J. Glucose metabolism in Neisseria gonorrhoeae. J Bacteriol. 1974 Nov;120(2):702–714. doi: 10.1128/jb.120.2.702-714.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mulks M. H., Knapp J. S. Immunoglobulin A1 protease types of Neisseria gonorrhoeae and their relationship to auxotype and serovar. Infect Immun. 1987 Apr;55(4):931–936. doi: 10.1128/iai.55.4.931-936.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mulks M. H., Kornfeld S. J., Frangione B., Plaut A. G. Relationship between the specificity of IgA proteases and serotypes in Haemophilus influenzae. J Infect Dis. 1982 Aug;146(2):266–274. doi: 10.1093/infdis/146.2.266. [DOI] [PubMed] [Google Scholar]
  16. Mulks M. H., Plaut A. G., Feldman H. A., Frangione B. IgA proteases of two distinct specificities are released by Neisseria meningitidis. J Exp Med. 1980 Nov 1;152(5):1442–1447. doi: 10.1084/jem.152.5.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mulks M. H., Plaut A. G. IgA protease production as a characteristic distinguishing pathogenic from harmless neisseriaceae. N Engl J Med. 1978 Nov 2;299(18):973–976. doi: 10.1056/NEJM197811022991802. [DOI] [PubMed] [Google Scholar]
  18. Plaut A. G. The IgA1 proteases of pathogenic bacteria. Annu Rev Microbiol. 1983;37:603–622. doi: 10.1146/annurev.mi.37.100183.003131. [DOI] [PubMed] [Google Scholar]
  19. Pohlner J., Halter R., Beyreuther K., Meyer T. F. Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease. 1987 Jan 29-Feb 4Nature. 325(6103):458–462. doi: 10.1038/325458a0. [DOI] [PubMed] [Google Scholar]

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