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. 1992 Feb;174(4):1352–1359. doi: 10.1128/jb.174.4.1352-1359.1992

The 14,000-molecular-weight antigen of Mycobacterium tuberculosis is related to the alpha-crystallin family of low-molecular-weight heat shock proteins.

A Verbon 1, R A Hartskeerl 1, A Schuitema 1, A H Kolk 1, D B Young 1, R Lathigra 1
PMCID: PMC206432  PMID: 1370952

Abstract

Eight monoclonal antibodies (MAbs) directed against the 14,000-molecular-weight (14K) antigen of Mycobacterium tuberculosis reacted specifically with mycobacteria of the M. tuberculosis complex. The nucleotide sequence of the gene encoding the 14K antigen was determined by using recombinant DNA clones isolated from lambda gt11 and cosmid libraries of the M. tuberculosis genome. The DNA sequence of the 14K protein gene coded for a polypeptide of 144 amino acids with a calculated molecular mass of 16,277 Da. The 14K antigen has a marked homology with proteins belonging to the alpha-crystallin family of low-molecular-weight heat shock proteins, which includes the 18K antigen of M. leprae. The eight MAbs recognized at least four distinct epitopes localized within the following three regions of the 14K protein: amino acids 10 to 92 (MAbs F67-8 and F67-16), amino acids 41 to 92 (F159-1 and F159-11), and amino acids 41 to 144 (F23-41, F24-2, F23-49, and TB68).

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Selected References

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

  1. 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]
  2. Baird P. N., Hall L. M., Coates A. R. A major antigen from Mycobacterium tuberculosis which is homologous to the heat shock proteins groES from E. coli and the htpA gene product of Coxiella burneti. Nucleic Acids Res. 1988 Sep 26;16(18):9047–9047. doi: 10.1093/nar/16.18.9047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Booth R. J., Harris D. P., Love J. M., Watson J. D. Antigenic proteins of Mycobacterium leprae. Complete sequence of the gene for the 18-kDa protein. J Immunol. 1988 Jan 15;140(2):597–601. [PubMed] [Google Scholar]
  5. Bossier P., Fitch I. T., Boucherie H., Tuite M. F. Structure and expression of a yeast gene encoding the small heat-shock protein Hsp26. Gene. 1989 May 30;78(2):323–330. doi: 10.1016/0378-1119(89)90234-5. [DOI] [PubMed] [Google Scholar]
  6. Calos M. P., Lebkowski J. S., Botchan M. R. High mutation frequency in DNA transfected into mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3015–3019. doi: 10.1073/pnas.80.10.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chandramuki A., Bothamley G. H., Brennan P. J., Ivanyi J. Levels of antibody to defined antigens of Mycobacterium tuberculosis in tuberculous meningitis. J Clin Microbiol. 1989 May;27(5):821–825. doi: 10.1128/jcm.27.5.821-825.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coates A. R., Hewitt J., Allen B. W., Ivanyi J., Mitchison D. A. Antigenic diversity of Mycobacterium tuberculosis and Mycobacterium bovis detected by means of monoclonal antibodies. Lancet. 1981 Jul 25;2(8239):167–169. doi: 10.1016/s0140-6736(81)90355-x. [DOI] [PubMed] [Google Scholar]
  9. Corpet F. Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res. 1988 Nov 25;16(22):10881–10890. doi: 10.1093/nar/16.22.10881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Falla J. C., Parra C. A., Mendoza M., Franco L. C., Guzmán F., Forero J., Orozco O., Patarroyo M. E. Identification of B- and T-cell epitopes within the MTP40 protein of Mycobacterium tuberculosis and their correlation with the disease course. Infect Immun. 1991 Jul;59(7):2265–2273. doi: 10.1128/iai.59.7.2265-2273.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garsia R. J., Hellqvist L., Booth R. J., Radford A. J., Britton W. J., Astbury L., Trent R. J., Basten A. Homology of the 70-kilodalton antigens from Mycobacterium leprae and Mycobacterium bovis with the Mycobacterium tuberculosis 71-kilodalton antigen and with the conserved heat shock protein 70 of eucaryotes. Infect Immun. 1989 Jan;57(1):204–212. doi: 10.1128/iai.57.1.204-212.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grimm B., Ish-Shalom D., Even D., Glaczinski H., Ottersbach P., Ohad I., Kloppstech K. The nuclear-coded chloroplast 22-kDa heat-shock protein of Chlamydomonas. Evidence for translocation into the organelle without a processing step. Eur J Biochem. 1989 Jul 1;182(3):539–546. doi: 10.1111/j.1432-1033.1989.tb14861.x. [DOI] [PubMed] [Google Scholar]
  14. Hartskeerl R. A., de Wit M. Y., Klatser P. R. Polymerase chain reaction for the detection of Mycobacterium leprae. J Gen Microbiol. 1989 Sep;135(9):2357–2364. doi: 10.1099/00221287-135-9-2357. [DOI] [PubMed] [Google Scholar]
  15. Hartskeerl R. A., van Rens R. M., Stabel L. F., de Wit M. Y., Klatser P. R. Selection and characterization of recombinant clones that produce Mycobacterium leprae antigens recognized by antibodies in sera from household contacts of leprosy patients. Infect Immun. 1990 Sep;58(9):2821–2827. doi: 10.1128/iai.58.9.2821-2827.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hartskeerl R., Tommassen J., Hoekstra W. Relationship between the proteins encoded by the exclusion determining locus of the IncI plasmid R144 and the cellular localization of these proteins in Escherichia coli K-12. Mol Gen Genet. 1985;200(1):138–144. doi: 10.1007/BF00383327. [DOI] [PubMed] [Google Scholar]
  17. Hickey E., Brandon S. E., Potter R., Stein G., Stein J., Weber L. A. Sequence and organization of genes encoding the human 27 kDa heat shock protein. Nucleic Acids Res. 1986 May 27;14(10):4127–4145. doi: 10.1093/nar/14.10.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hunter S. W., Rivoire B., Mehra V., Bloom B. R., Brennan P. J. The major native proteins of the leprosy bacillus. J Biol Chem. 1990 Aug 25;265(24):14065–14068. [PubMed] [Google Scholar]
  19. Ingolia T. D., Craig E. A. Four small Drosophila heat shock proteins are related to each other and to mammalian alpha-crystallin. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2360–2364. doi: 10.1073/pnas.79.7.2360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ivanyi J., Bothamley G. H., Jackett P. S. Immunodiagnostic assays for tuberculosis and leprosy. Br Med Bull. 1988 Jul;44(3):635–649. doi: 10.1093/oxfordjournals.bmb.a072273. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Kingston A. E., Salgame P. R., Mitchison N. A., Colston M. J. Immunological activity of a 14-kilodalton recombinant protein of Mycobacterium tuberculosis H37Rv. Infect Immun. 1987 Dec;55(12):3149–3154. doi: 10.1128/iai.55.12.3149-3154.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kolk A. H., Evers R., Groothuis D. G., Gilis H., Kuijper S. Production and characterization of monoclonal antibodies against specific serotypes of Mycobacterium avium and the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum complex. Infect Immun. 1989 Aug;57(8):2514–2521. doi: 10.1128/iai.57.8.2514-2521.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kolk A. H., Ho M. L., Klatser P. R., Eggelte T. A., Kuijper S., de Jonge S., van Leeuwen J. Production and characterization of monoclonal antibodies to Mycobacterium tuberculosis, M. bovis (BCG) and M. leprae. Clin Exp Immunol. 1984 Dec;58(3):511–521. [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Matthews R., Scoging A., Rees A. D. Mycobacterial antigen-specific human T-cell clones secreting macrophage activating factors. Immunology. 1985 Jan;54(1):17–23. [PMC free article] [PubMed] [Google Scholar]
  27. McElwain E. F., Spiker S. A wheat cDNA clone which is homologous to the 17 kd heat-shock protein gene family of soybean. Nucleic Acids Res. 1989 Feb 25;17(4):1764–1764. doi: 10.1093/nar/17.4.1764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mehra V., Sweetser D., Young R. A. Efficient mapping of protein antigenic determinants. Proc Natl Acad Sci U S A. 1986 Sep;83(18):7013–7017. doi: 10.1073/pnas.83.18.7013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Murray C. J., Styblo K., Rouillon A. Tuberculosis in developing countries: burden, intervention and cost. Bull Int Union Tuberc Lung Dis. 1990 Mar;65(1):6–24. [PubMed] [Google Scholar]
  30. Nerland A. H., Mustafa A. S., Sweetser D., Godal T., Young R. A. A protein antigen of Mycobacterium leprae is related to a family of small heat shock proteins. J Bacteriol. 1988 Dec;170(12):5919–5921. doi: 10.1128/jb.170.12.5919-5921.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Raschke E., Baumann G., Schöffl F. Nucleotide sequence analysis of soybean small heat shock protein genes belonging to two different multigene families. J Mol Biol. 1988 Feb 20;199(4):549–557. doi: 10.1016/0022-2836(88)90300-2. [DOI] [PubMed] [Google Scholar]
  32. Results of a World Health Organization-sponsored workshop on monoclonal antibodies to Mycobacterium leprae. Infect Immun. 1985 May;48(2):603–605. doi: 10.1128/iai.48.2.603-605.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Results of a World Health Organization-sponsored workshop to characterize antigens recognized by mycobacterium-specific monoclonal antibodies. Infect Immun. 1986 Feb;51(2):718–720. doi: 10.1128/iai.51.2.718-720.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schöningh R., Verstijnen C. P., Kuijper S., Kolk A. H. Enzyme immunoassay for identification of heat-killed mycobacteria belonging to the Mycobacterium tuberculosis and Mycobacterium avium complexes and derived from early cultures. J Clin Microbiol. 1990 Apr;28(4):708–713. doi: 10.1128/jcm.28.4.708-713.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shinnick T. M., Krat C., Schadow S. Isolation and restriction site maps of the genes encoding five Mycobacterium tuberculosis proteins. Infect Immun. 1987 Jul;55(7):1718–1721. doi: 10.1128/iai.55.7.1718-1721.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shinnick T. M. The 65-kilodalton antigen of Mycobacterium tuberculosis. J Bacteriol. 1987 Mar;169(3):1080–1088. doi: 10.1128/jb.169.3.1080-1088.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shinnick T. M., Vodkin M. H., Williams J. C. The Mycobacterium tuberculosis 65-kilodalton antigen is a heat shock protein which corresponds to common antigen and to the Escherichia coli GroEL protein. Infect Immun. 1988 Feb;56(2):446–451. doi: 10.1128/iai.56.2.446-451.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  40. Stanley K. K., Luzio J. P. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J. 1984 Jun;3(6):1429–1434. doi: 10.1002/j.1460-2075.1984.tb01988.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Susek R. E., Lindquist S. L. hsp26 of Saccharomyces cerevisiae is related to the superfamily of small heat shock proteins but is without a demonstrable function. Mol Cell Biol. 1989 Nov;9(11):5265–5271. doi: 10.1128/mcb.9.11.5265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Takahashi T., Komeda Y. Characterization of two genes encoding small heat-shock proteins in Arabidopsis thaliana. Mol Gen Genet. 1989 Nov;219(3):365–372. doi: 10.1007/BF00259608. [DOI] [PubMed] [Google Scholar]
  43. Van Eys G. J., Zaal J., Schoone G. J., Terpstra W. J. DNA hybridization with hardjobovis-specific recombinant probes as a method for type discrimination of Leptospira interrogans serovar hardjo. J Gen Microbiol. 1988 Mar;134(3):567–574. doi: 10.1099/00221287-134-3-567. [DOI] [PubMed] [Google Scholar]
  44. Verbon A., Kuijper S., Jansen H. M., Speelman P., Kolk A. H. Antigens in culture supernatant of Mycobacterium tuberculosis: epitopes defined by monoclonal and human antibodies. J Gen Microbiol. 1990 May;136(5):955–964. doi: 10.1099/00221287-136-5-955. [DOI] [PubMed] [Google Scholar]
  45. Verstijnen C. P., Schöningh R., Kuijper S., Bruins J., von Ketel R. J., Groothuis D. G., Kolk A. H. Rapid identification of cultured Mycobacterium tuberculosis with a panel of monoclonal antibodies in western blot and immunofluorescence. Res Microbiol. 1989 Nov-Dec;140(9):653–666. doi: 10.1016/0923-2508(89)90197-6. [DOI] [PubMed] [Google Scholar]
  46. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  47. Young D. B., Garbe T. R. Heat shock proteins and antigens of Mycobacterium tuberculosis. Infect Immun. 1991 Sep;59(9):3086–3093. doi: 10.1128/iai.59.9.3086-3093.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Young D. B., Kent L., Young R. A. Screening of a recombinant mycobacterial DNA library with polyclonal antiserum and molecular weight analysis of expressed antigens. Infect Immun. 1987 Jun;55(6):1421–1425. doi: 10.1128/iai.55.6.1421-1425.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Young D., Lathigra R., Hendrix R., Sweetser D., Young R. A. Stress proteins are immune targets in leprosy and tuberculosis. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4267–4270. doi: 10.1073/pnas.85.12.4267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Young R. A., Bloom B. R., Grosskinsky C. M., Ivanyi J., Thomas D., Davis R. W. Dissection of Mycobacterium tuberculosis antigens using recombinant DNA. Proc Natl Acad Sci U S A. 1985 May;82(9):2583–2587. doi: 10.1073/pnas.82.9.2583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Zabeau M., Stanley K. K. Enhanced expression of cro-beta-galactosidase fusion proteins under the control of the PR promoter of bacteriophage lambda. EMBO J. 1982;1(10):1217–1224. doi: 10.1002/j.1460-2075.1982.tb00016.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. de Jong W. W., Leunissen J. A., Leenen P. J., Zweers A., Versteeg M. Dogfish alpha-crystallin sequences. Comparison with small heat shock proteins and Schistosoma egg antigen. J Biol Chem. 1988 Apr 15;263(11):5141–5149. [PubMed] [Google Scholar]

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