Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Jun;59(6):1905–1910. doi: 10.1128/iai.59.6.1905-1910.1991

Proteins released from Mycobacterium tuberculosis during growth.

P Andersen 1, D Askgaard 1, L Ljungqvist 1, J Bennedsen 1, I Heron 1
PMCID: PMC257941  PMID: 1903768

Abstract

Proteins secreted from Mycobacterium tuberculosis during growth are believed to be important for protective immunity against tuberculosis. We have investigated the growth of M. tuberculosis in an enriched liquid medium. The release of isocitrate dehydrogenase from the bacilli served as a marker of autolysis and was observed during the late logarithmic growth phase. The release of proteins during the culture period was investigated by enzyme-linked immunosorbent assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Three major groups of proteins, which differed markedly with respect to profile of release and location in intact bacilli, were defined. A short-term filtrate devoid of autolytic products was defined and found to be composed of 33 major components. Five proteins were identified by monoclonal antibodies. Pronounced superoxide dismutase activity was detected in the filtrate. The enzyme was purified and identified as a dominating component of short-term filtrate.

Full text

PDF
1905

Images in this article

1907Image
on p.1907
1908Image
on p.1908
1909Image
on p.1909

Selected References

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

  1. Abou-Zeid C., Ratliff T. L., Wiker H. G., Harboe M., Bennedsen J., Rook G. A. Characterization of fibronectin-binding antigens released by Mycobacterium tuberculosis and Mycobacterium bovis BCG. Infect Immun. 1988 Dec;56(12):3046–3051. doi: 10.1128/iai.56.12.3046-3051.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abou-Zeid C., Smith I., Grange J. M., Ratliff T. L., Steele J., Rook G. A. The secreted antigens of Mycobacterium tuberculosis and their relationship to those recognized by the available antibodies. J Gen Microbiol. 1988 Feb;134(2):531–538. doi: 10.1099/00221287-134-2-531. [DOI] [PubMed] [Google Scholar]
  3. Andersen A. B., Yuan Z. L., Hasløv K., Vergmann B., Bennedsen J. Interspecies reactivity of five monoclonal antibodies to Mycobacterium tuberculosis as examined by immunoblotting and enzyme-linked immunosorbent assay. J Clin Microbiol. 1986 Mar;23(3):446–451. doi: 10.1128/jcm.23.3.446-451.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andersen P., Askgaard D., Ljungqvist L., Bentzon M. W., Heron I. T-cell proliferative response to antigens secreted by Mycobacterium tuberculosis. Infect Immun. 1991 Apr;59(4):1558–1563. doi: 10.1128/iai.59.4.1558-1563.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beauchamp C., Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971 Nov;44(1):276–287. doi: 10.1016/0003-2697(71)90370-8. [DOI] [PubMed] [Google Scholar]
  6. Borremans M., de Wit L., Volckaert G., Ooms J., de Bruyn J., Huygen K., van Vooren J. P., Stelandre M., Verhofstadt R., Content J. Cloning, sequence determination, and expression of a 32-kilodalton-protein gene of Mycobacterium tuberculosis. Infect Immun. 1989 Oct;57(10):3123–3130. doi: 10.1128/iai.57.10.3123-3130.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Collins F. M., Lamb J. R., Young D. B. Biological activity of protein antigens isolated from Mycobacterium tuberculosis culture filtrate. Infect Immun. 1988 May;56(5):1260–1266. doi: 10.1128/iai.56.5.1260-1266.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. David H. L., Lévy-Frébault V., Thorel M. F. Characterization of distinct layers of the Mycobacterium avium envelope in respect of their composition by fatty acids, proteins, oligosaccharides and antigens. Zentralbl Bakteriol Mikrobiol Hyg A. 1988 Apr;268(2):193–208. doi: 10.1016/s0176-6724(88)80003-8. [DOI] [PubMed] [Google Scholar]
  10. Harboe M., Nagai S., Patarroyo M. E., Torres M. L., Ramirez C., Cruz N. Properties of proteins MPB64, MPB70, and MPB80 of Mycobacterium bovis BCG. Infect Immun. 1986 Apr;52(1):293–302. doi: 10.1128/iai.52.1.293-302.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kusunose E., Ichihara K., Noda Y., Kusunose M. Superoxide dismutase from Mycobacterium tuberculosis. J Biochem. 1976 Dec;80(6):1343–1352. doi: 10.1093/oxfordjournals.jbchem.a131407. [DOI] [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. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  14. Nagai S., Wiker H. G., Harboe M., Kinomoto M. Isolation and partial characterization of major protein antigens in the culture fluid of Mycobacterium tuberculosis. Infect Immun. 1991 Jan;59(1):372–382. doi: 10.1128/iai.59.1.372-382.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Oliver D. Protein secretion in Escherichia coli. Annu Rev Microbiol. 1985;39:615–648. doi: 10.1146/annurev.mi.39.100185.003151. [DOI] [PubMed] [Google Scholar]
  16. Orme I. M. Induction of nonspecific acquired resistance and delayed-type hypersensitivity, but not specific acquired resistance in mice inoculated with killed mycobacterial vaccines. Infect Immun. 1988 Dec;56(12):3310–3312. doi: 10.1128/iai.56.12.3310-3312.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Prioli R. P., Tanna A., Brown I. N. Rapid methods for counting mycobacteria--comparison of methods for extraction of mycobacterial adenosine triphosphate (ATP) determined by firefly luciferase assay. Tubercle. 1985 Jun;66(2):99–108. doi: 10.1016/0041-3879(85)90074-1. [DOI] [PubMed] [Google Scholar]
  18. Thangaraj H. S., Lamb F. I., Davis E. O., Jenner P. J., Jeyakumar L. H., Colston M. J. Identification, sequencing, and expression of Mycobacterium leprae superoxide dismutase, a major antigen. Infect Immun. 1990 Jun;58(6):1937–1942. doi: 10.1128/iai.58.6.1937-1942.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Watson J. D. Leprosy: understanding protective immunity. Immunol Today. 1989 Jul;10(7):218–221. doi: 10.1016/0167-5699(89)90253-3. [DOI] [PubMed] [Google Scholar]
  20. Wiker H. G., Harboe M., Nagai S., Bennedsen J. Quantitative and qualitative studies on the major extracellular antigen of Mycobacterium tuberculosis H37Rv and Mycobacterium bovis BCG. Am Rev Respir Dis. 1990 Apr;141(4 Pt 1):830–838. doi: 10.1164/ajrccm/141.4_Pt_1.830. [DOI] [PubMed] [Google Scholar]
  21. Wiker H. G., Sletten K., Nagai S., Harboe M. Evidence for three separate genes encoding the proteins of the mycobacterial antigen 85 complex. Infect Immun. 1990 Jan;58(1):272–274. doi: 10.1128/iai.58.1.272-274.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Worsaae A., Ljungqvist L., Hasløv K., Heron I., Bennedsen J. Allergenic and blastogenic reactivity of three antigens from Mycobacterium tuberculosis in sensitized guinea pigs. Infect Immun. 1987 Dec;55(12):2922–2927. doi: 10.1128/iai.55.12.2922-2927.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Worsaae A., Ljungqvist L., Heron I. Monoclonal antibodies produced in BALB.B10 mice define new antigenic determinants in culture filtrate preparations of Mycobacterium tuberculosis. J Clin Microbiol. 1988 Dec;26(12):2608–2614. doi: 10.1128/jcm.26.12.2608-2614.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yamaguchi R., Matsuo K., Yamazaki A., Abe C., Nagai S., Terasaka K., Yamada T. Cloning and characterization of the gene for immunogenic protein MPB64 of Mycobacterium bovis BCG. Infect Immun. 1989 Jan;57(1):283–288. doi: 10.1128/iai.57.1.283-288.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES