Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1995 Feb;63(2):580–584. doi: 10.1128/iai.63.2.580-584.1995

Antigenic and structural similarities between Mycobacterium tuberculosis 50- to 55-kilodalton and Mycobacterium bovis BCG 45- to 47-kilodalton antigens.

C Espitia 1, R Espinosa 1, R Saavedra 1, R Mancilla 1, F Romain 1, A Laqueyrerie 1, C Moreno 1
PMCID: PMC173035  PMID: 7822025

Abstract

The relationship between Mycobacterium tuberculosis 50- to 55-kDa protein and Mycobacterium bovis BCG 45- to 47-kDa antigen was examined by using immunological and biochemical criteria. Reciprocal cross-reactivity with a rabbit polyclonal antiserum against the M. bovis BCG protein and with a monoclonal antibody raised against the M. tuberculosis antigen was observed. The epitope recognized by this antibody was apparently present only in proteins of M. tuberculosis and M. bovis BCG among the 11 mycobacterial species tested. The amino-terminal sequences and total amino acid contents of these proteins showed strong similarities. Both antigens are glycoproteins as assessed by binding of concanavalin A, labeling of carbohydrate moieties with biotin-hydrazide, and digestion of carbohydrates with jack bean alpha-D-mannosidase, which produced a reduction of the molecular weights of the proteins and totally eliminated concanavalin A binding. Both M. tuberculosis and M. bovis BCG proteins are secreted, since they were found mainly in the culture medium. Analysis of M. tuberculosis 50- to 55-kDa antigen by two-dimensional gel electrophoresis showed at least seven different components, as previously described for the M. bovis BCG antigen. Solid-phase immunoassays showed that the purified M. tuberculosis 50- to 55-kDa protein was recognized by serum specimens from 70% of individuals with pulmonary tuberculosis from a total of 77 Mexican patients examined.

Full Text

The Full Text of this article is available as a PDF (268.3 KB).

Selected References

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

  1. Espitia C., Cervera I., González R., Mancilla R. A 38-kD Mycobacterium tuberculosis antigen associated with infection. Its isolation and serologic evaluation. Clin Exp Immunol. 1989 Sep;77(3):373–377. [PMC free article] [PubMed] [Google Scholar]
  2. Espitia C., Mancilla R. Identification, isolation and partial characterization of Mycobacterium tuberculosis glycoprotein antigens. Clin Exp Immunol. 1989 Sep;77(3):378–383. [PMC free article] [PubMed] [Google Scholar]
  3. Espitia C., Sciutto E., Bottasso O., González-Amaro R., Hernández-Pando R., Mancilla R. High antibody levels to the mycobacterial fibronectin-binding antigen of 30-31 kD in tuberculosis and lepromatous leprosy. Clin Exp Immunol. 1992 Mar;87(3):362–367. doi: 10.1111/j.1365-2249.1992.tb03003.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fifis T., Costopoulos C., Radford A. J., Bacic A., Wood P. R. Purification and characterization of major antigens from a Mycobacterium bovis culture filtrate. Infect Immun. 1991 Mar;59(3):800–807. doi: 10.1128/iai.59.3.800-807.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Garbe T., Harris D., Vordermeier M., Lathigra R., Ivanyi J., Young D. Expression of the Mycobacterium tuberculosis 19-kilodalton antigen in Mycobacterium smegmatis: immunological analysis and evidence of glycosylation. Infect Immun. 1993 Jan;61(1):260–267. doi: 10.1128/iai.61.1.260-267.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hardy M. R., Townsend R. R., Lee Y. C. Monosaccharide analysis of glycoconjugates by anion exchange chromatography with pulsed amperometric detection. Anal Biochem. 1988 Apr;170(1):54–62. doi: 10.1016/0003-2697(88)90089-9. [DOI] [PubMed] [Google Scholar]
  7. Harris D. P., Vordermeier H. M., Roman E., Lathigra R., Brett S. J., Moreno C., Ivanyi J. Murine T cell-stimulatory peptides from the 19-kDa antigen of Mycobacterium tuberculosis. Epitope-restricted homology with the 28-kDa protein of Mycobacterium leprae. J Immunol. 1991 Oct 15;147(8):2706–2712. [PubMed] [Google Scholar]
  8. Jackett P. S., Bothamley G. H., Batra H. V., Mistry A., Young D. B., Ivanyi J. Specificity of antibodies to immunodominant mycobacterial antigens in pulmonary tuberculosis. J Clin Microbiol. 1988 Nov;26(11):2313–2318. doi: 10.1128/jcm.26.11.2313-2318.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. O'Shannessy D. J. Antibodies biotinylated via sugar moieties. Methods Enzymol. 1990;184:162–166. doi: 10.1016/0076-6879(90)84270-q. [DOI] [PubMed] [Google Scholar]
  14. Orme I. M., Andersen P., Boom W. H. T cell response to Mycobacterium tuberculosis. J Infect Dis. 1993 Jun;167(6):1481–1497. doi: 10.1093/infdis/167.6.1481. [DOI] [PubMed] [Google Scholar]
  15. Pal P. G., Horwitz M. A. Immunization with extracellular proteins of Mycobacterium tuberculosis induces cell-mediated immune responses and substantial protective immunity in a guinea pig model of pulmonary tuberculosis. Infect Immun. 1992 Nov;60(11):4781–4792. doi: 10.1128/iai.60.11.4781-4792.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Parekh B. S., Mehta H. B., West M. D., Montelaro R. C. Preparative elution of proteins from nitrocellulose membranes after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Anal Biochem. 1985 Jul;148(1):87–92. doi: 10.1016/0003-2697(85)90631-1. [DOI] [PubMed] [Google Scholar]
  17. Romain F., Augier J., Pescher P., Marchal G. Isolation of a proline-rich mycobacterial protein eliciting delayed-type hypersensitivity reactions only in guinea pigs immunized with living mycobacteria. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5322–5326. doi: 10.1073/pnas.90.11.5322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Romain F., Laqueyrerie A., Militzer P., Pescher P., Chavarot P., Lagranderie M., Auregan G., Gheorghiu M., Marchal G. Identification of a Mycobacterium bovis BCG 45/47-kilodalton antigen complex, an immunodominant target for antibody response after immunization with living bacteria. Infect Immun. 1993 Feb;61(2):742–750. doi: 10.1128/iai.61.2.742-750.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vordermeier H. M., Harris D. P., Friscia G., Román E., Surcel H. M., Moreno C., Pasvol G., Ivanyi J. T cell repertoire in tuberculosis: selective anergy to an immunodominant epitope of the 38-kDa antigen in patients with active disease. Eur J Immunol. 1992 Oct;22(10):2631–2637. doi: 10.1002/eji.1830221024. [DOI] [PubMed] [Google Scholar]
  21. Wieles B., van Agterveld M., Janson A., Clark-Curtiss J., Rinke de Wit T., Harboe M., Thole J. Characterization of a Mycobacterium leprae antigen related to the secreted Mycobacterium tuberculosis protein MPT32. Infect Immun. 1994 Jan;62(1):252–258. doi: 10.1128/iai.62.1.252-258.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wiker H. G., Harboe M. The antigen 85 complex: a major secretion product of Mycobacterium tuberculosis. Microbiol Rev. 1992 Dec;56(4):648–661. doi: 10.1128/mr.56.4.648-661.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES