Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1994 Jul 1;180(1):319–327. doi: 10.1084/jem.180.1.319

Purification, characterization, gene sequence, and significance of a bacterioferritin from Mycobacterium leprae

PMCID: PMC2191570  PMID: 8006590

Abstract

The study of tissue-derived Mycobacterium leprae provides insights to the immunopathology of leprosy and helps identify broad molecular features necessary for mycobacterial parasitism. A major membrane protein (MMP-II) of in vivo-derived M. leprae previously recognized (Hunter, S.W., B. Rivoire, V. Mehra, B.R. Bloom, and P.J. Brennan. 1990. J. Biol. Chem. 265:14065) was purified from extracts of the organism and partial amino acid sequence obtained. This information allowed recognition, within one of the cosmids that encompass the entire M. leprae genome, of a complete gene, bfr, encoding a protein of subunit size 18.2 kD. The amino acid sequence deduced from the major membrane protein II (MMP-II) gene revealed considerable homology to several bacterioferritins. Analysis of the native protein demonstrated the iron content, absorption spectrum, and large native molecular mass (380 kD) of several known bacterioferritins. The ferroxidase-center residues typical of ferritins were conserved in the M. leprae product. Oligonucleotides derived from the amino acid sequence of M. leprae bacterioferritin enabled amplification of much of the MMP-II gene and the detection of homologous sequences in Mycobacterium paratuberculosis, Mycobacterium avium, Mycobacterium tuberculosis, Mycobacterium intracellulare, and Mycobacterium scrofulaceum. The role of this iron-rich protein in the virulence of M. leprae is discussed.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Andrews S. C., Harrison P. M., Guest J. R. Cloning, sequencing, and mapping of the bacterioferritin gene (bfr) of Escherichia coli K-12. J Bacteriol. 1989 Jul;171(7):3940–3947. doi: 10.1128/jb.171.7.3940-3947.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bauminger E. R., Cohen S. G., Dickson D. P., Levy A., Ofer S., Yariv J. Mössbauer spectroscopy of Escherichia coli and its iron-storage protein. Biochim Biophys Acta. 1980 Jun 26;623(2):237–242. doi: 10.1016/0005-2795(80)90252-4. [DOI] [PubMed] [Google Scholar]
  3. Brooks B. W., Young N. M., Watson D. C., Robertson R. H., Sugden E. A., Nielsen K. H., Becker S. A. Mycobacterium paratuberculosis antigen D: characterization and evidence that it is a bacterioferritin. J Clin Microbiol. 1991 Aug;29(8):1652–1658. doi: 10.1128/jcm.29.8.1652-1658.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cheesman M. R., Thomson A. J., Greenwood C., Moore G. R., Kadir F. Bis-methionine axial ligation of haem in bacterioferritin from Pseudomonas aeruginosa. Nature. 1990 Aug 23;346(6286):771–773. doi: 10.1038/346771a0. [DOI] [PubMed] [Google Scholar]
  5. Church G. M., Kieffer-Higgins S. Multiplex DNA sequencing. Science. 1988 Apr 8;240(4849):185–188. doi: 10.1126/science.3353714. [DOI] [PubMed] [Google Scholar]
  6. Davis M. T., Lee T. D. Analysis of peptide mixtures by capillary high performance liquid chromatography: a practical guide to small-scale separations. Protein Sci. 1992 Jul;1(7):935–944. doi: 10.1002/pro.5560010712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eiglmeier K., Honoré N., Woods S. A., Caudron B., Cole S. T. Use of an ordered cosmid library to deduce the genomic organization of Mycobacterium leprae. Mol Microbiol. 1993 Jan;7(2):197–206. doi: 10.1111/j.1365-2958.1993.tb01111.x. [DOI] [PubMed] [Google Scholar]
  8. Finch C. A., Huebers H. Perspectives in iron metabolism. N Engl J Med. 1982 Jun 24;306(25):1520–1528. doi: 10.1056/NEJM198206243062504. [DOI] [PubMed] [Google Scholar]
  9. Grossman M. J., Hinton S. M., Minak-Bernero V., Slaughter C., Stiefel E. I. Unification of the ferritin family of proteins. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2419–2423. doi: 10.1073/pnas.89.6.2419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hall R. M., Sritharan M., Messenger A. J., Ratledge C. Iron transport in Mycobacterium smegmatis: occurrence of iron-regulated envelope proteins as potential receptors for iron uptake. J Gen Microbiol. 1987 Aug;133(8):2107–2114. doi: 10.1099/00221287-133-8-2107. [DOI] [PubMed] [Google Scholar]
  11. Harrison R. F., Hubscher S. G. The spectrum of bile duct lesions in end-stage primary sclerosing cholangitis. Histopathology. 1991 Oct;19(4):321–327. doi: 10.1111/j.1365-2559.1991.tb00046.x. [DOI] [PubMed] [Google Scholar]
  12. Hawke D. H., Harris D. C., Shively J. E. Microsequence analysis of peptides and proteins. V. Design and performance of a novel gas-liquid-solid phase instrument. Anal Biochem. 1985 Jun;147(2):315–330. doi: 10.1016/0003-2697(85)90278-7. [DOI] [PubMed] [Google Scholar]
  13. Honoré N., Bergh S., Chanteau S., Doucet-Populaire F., Eiglmeier K., Garnier T., Georges C., Launois P., Limpaiboon T., Newton S. Nucleotide sequence of the first cosmid from the Mycobacterium leprae genome project: structure and function of the Rif-Str regions. Mol Microbiol. 1993 Jan;7(2):207–214. doi: 10.1111/j.1365-2958.1993.tb01112.x. [DOI] [PubMed] [Google Scholar]
  14. Hunter S. W., Fujiwara T., Brennan P. J. Structure and antigenicity of the major specific glycolipid antigen of Mycobacterium leprae. J Biol Chem. 1982 Dec 25;257(24):15072–15078. [PubMed] [Google Scholar]
  15. 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]
  16. Jacobs W. R., Jr, Kalpana G. V., Cirillo J. D., Pascopella L., Snapper S. B., Udani R. A., Jones W., Barletta R. G., Bloom B. R. Genetic systems for mycobacteria. Methods Enzymol. 1991;204:537–555. doi: 10.1016/0076-6879(91)04027-l. [DOI] [PubMed] [Google Scholar]
  17. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Laulhère J. P., Labouré A. M., Van Wuytswinkel O., Gagnon J., Briat J. F. Purification, characterization and function of bacterioferritin from the cyanobacterium Synechocystis P.C.C. 6803. Biochem J. 1992 Feb 1;281(Pt 3):785–793. doi: 10.1042/bj2810785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lee B. Y., Hefta S. A., Brennan P. J. Characterization of the major membrane protein of virulent Mycobacterium tuberculosis. Infect Immun. 1992 May;60(5):2066–2074. doi: 10.1128/iai.60.5.2066-2074.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lee T. D., Shively J. E. Enzymatic and chemical digestion of proteins for mass spectrometry. Methods Enzymol. 1990;193:361–374. doi: 10.1016/0076-6879(90)93427-m. [DOI] [PubMed] [Google Scholar]
  22. Liesack W., Pitulle C., Sela S., Stackebrandt E. Nucleotide sequence of the 16S rRNA from Mycobacterium leprae. Nucleic Acids Res. 1990 Sep 25;18(18):5558–5558. doi: 10.1093/nar/18.18.5558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Litwin C. M., Calderwood S. B. Role of iron in regulation of virulence genes. Clin Microbiol Rev. 1993 Apr;6(2):137–149. doi: 10.1128/cmr.6.2.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Matzanke B. F., Müller G. I., Bill E., Trautwein A. X. Iron metabolism of Escherichia coli studied by Mössbauer spectroscopy and biochemical methods. Eur J Biochem. 1989 Aug 1;183(2):371–379. doi: 10.1111/j.1432-1033.1989.tb14938.x. [DOI] [PubMed] [Google Scholar]
  25. Merkal R. S., Curran B. J. Growth and metabolic characteristics of Mycobacterium paratuberculosis. Appl Microbiol. 1974 Aug;28(2):276–279. doi: 10.1128/am.28.2.276-279.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moore G. R., Mann S., Bannister J. V. Isolation and properties of the complex nonheme-iron-containing cytochrome b557 (bacterioferritin) from Pseudomonas aeruginosa. J Inorg Biochem. 1986 Oct-Nov;28(2-3):329–336. doi: 10.1016/0162-0134(86)80097-6. [DOI] [PubMed] [Google Scholar]
  27. Noordeen S. K., Lopez Bravo L., Sundaresan T. K. Estimated number of leprosy cases in the world. Bull World Health Organ. 1992;70(1):7–10. [PMC free article] [PubMed] [Google Scholar]
  28. Shively J. E., Miller P., Ronk M. Microsequence analysis of peptides and proteins. VI. A continuous flow reactor for sample concentration and sequence analysis. Anal Biochem. 1987 Jun;163(2):517–529. doi: 10.1016/0003-2697(87)90257-0. [DOI] [PubMed] [Google Scholar]
  29. Smida J., Kazda J., Stackebrandt E. Molecular-genetic evidence for the relationship of Mycobacterium leprae to slow-growing pathogenic mycobacteria. Int J Lepr Other Mycobact Dis. 1988 Sep;56(3):449–454. [PubMed] [Google Scholar]
  30. Smith D. R., Calvo J. M. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase. Nucleic Acids Res. 1980 May 24;8(10):2255–2274. doi: 10.1093/nar/8.10.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Smith J. M., Ford G. C., Harrison P. M., Yariv J., Kalb A. J. Molecular size and symmetry of the bacterioferritin of Escherichia coli. X-ray crystallographic characterization of four crystal forms. J Mol Biol. 1989 Jan 20;205(2):465–467. doi: 10.1016/0022-2836(89)90358-6. [DOI] [PubMed] [Google Scholar]
  32. Smith J. M., Quirk A. V., Plank R. W., Diffin F. M., Ford G. C., Harrison P. M. The identity of Escherichia coli bacterioferritin and cytochrome b1. Biochem J. 1988 Oct 15;255(2):737–740. [PMC free article] [PubMed] [Google Scholar]
  33. Sugden E. A., Brooks B. W., Young N. M., Watson D. C., Nielsen K. H., Corner A. H., Turcotte C., Michaelides A., Stewart R. B. Chromatographic purification and characterization of antigens A and D from Mycobacterium paratuberculosis and their use in enzyme-linked immunosorbent assays for diagnosis of paratuberculosis in sheep. J Clin Microbiol. 1991 Aug;29(8):1659–1664. doi: 10.1128/jcm.29.8.1659-1664.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Theil E. C. Ferritin: structure, gene regulation, and cellular function in animals, plants, and microorganisms. Annu Rev Biochem. 1987;56:289–315. doi: 10.1146/annurev.bi.56.070187.001445. [DOI] [PubMed] [Google Scholar]
  35. Titani K., Sasagawa T., Resing K., Walsh K. A. A simple and rapid purification of commercial trypsin and chymotrypsin by reverse-phase high-performance liquid chromatography. Anal Biochem. 1982 Jul 1;123(2):408–412. doi: 10.1016/0003-2697(82)90465-1. [DOI] [PubMed] [Google Scholar]
  36. Wheeler P. R. Recent research into the physiology of Mycobacterium leprae. Adv Microb Physiol. 1990;31:71–124. doi: 10.1016/s0065-2911(08)60120-4. [DOI] [PubMed] [Google Scholar]
  37. Yariv J., Kalb A. J., Sperling R., Bauminger E. R., Cohen S. G., Ofer S. The composition and the structure of bacterioferritin of Escherichia coli. Biochem J. 1981 Jul 1;197(1):171–175. doi: 10.1042/bj1970171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Young D. B., Kaufmann S. H., Hermans P. W., Thole J. E. Mycobacterial protein antigens: a compilation. Mol Microbiol. 1992 Jan;6(2):133–145. doi: 10.1111/j.1365-2958.1992.tb01994.x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES