Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2001 Oct 1;359(Pt 1):17–22. doi: 10.1042/0264-6021:3590017

Lipid modification of the Cu,Zn superoxide dismutase from Mycobacterium tuberculosis.

M D'orazio 1, S Folcarelli 1, F Mariani 1, V Colizzi 1, G Rotilio 1, A Battistoni 1
PMCID: PMC1222117  PMID: 11563965

Abstract

The leader sequence of Mycobacterium tuberculosis Cu,Zn superoxide dismutase (Cu,ZnSOD) contains a prokaryotic membrane lipoprotein attachment site. In the present study, we have found that the protein, which exhibits detectable SOD activity, is lipid-modified and associated with the bacterial membrane when expressed either in M. tuberculosis or in Escherichia coli. These results provide the first demonstration of lipid modification of a Cu,ZnSOD. An analysis of the sodC genes present in available databases indicates that the same signal for lipid modification is also present in the sodC gene products from other mycobacteria and Gram-positive bacteria and, uniquely, in two distinct sodC gene products from the Gram-negative bacterium Salmonella typhimurium. Evidence is also provided for an up-regulation of M. tuberculosis sodC in response to phagocytosis by human macrophages, suggesting that Cu,ZnSOD is involved in the mechanisms that facilitate mycobacterial intracellular growth.

Full Text

The Full Text of this article is available as a PDF (155.8 KB).

Selected References

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

  1. Battistoni A., Donnarumma G., Greco R., Valenti P., Rotilio G. Overexpression of a hydrogen peroxide-resistant periplasmic Cu,Zn superoxide dismutase protects Escherichia coli from macrophage killing. Biochem Biophys Res Commun. 1998 Feb 24;243(3):804–807. doi: 10.1006/bbrc.1998.8182. [DOI] [PubMed] [Google Scholar]
  2. Battistoni A., Pacello F., Folcarelli S., Ajello M., Donnarumma G., Greco R., Ammendolia M. G., Touati D., Rotilio G., Valenti P. Increased expression of periplasmic Cu,Zn superoxide dismutase enhances survival of Escherichia coli invasive strains within nonphagocytic cells. Infect Immun. 2000 Jan;68(1):30–37. doi: 10.1128/iai.68.1.30-37.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Battistoni A., Rotilio G. Isolation of an active and heat-stable monomeric form of Cu,Zn superoxide dismutase from the periplasmic space of Escherichia coli. FEBS Lett. 1995 Oct 30;374(2):199–202. doi: 10.1016/0014-5793(95)01106-o. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Bordier C. Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981 Feb 25;256(4):1604–1607. [PubMed] [Google Scholar]
  6. Bordo D., Djinović K., Bolognesi M. Conserved patterns in the Cu,Zn superoxide dismutase family. J Mol Biol. 1994 May 6;238(3):366–386. doi: 10.1006/jmbi.1994.1298. [DOI] [PubMed] [Google Scholar]
  7. Bordo D., Matak D., Djinovic-Carugo K., Rosano C., Pesce A., Bolognesi M., Stroppolo M. E., Falconi M., Battistoni A., Desideri A. Evolutionary constraints for dimer formation in prokaryotic Cu,Zn superoxide dismutase. J Mol Biol. 1999 Jan 8;285(1):283–296. doi: 10.1006/jmbi.1998.2267. [DOI] [PubMed] [Google Scholar]
  8. Carlioz A., Touati D. Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J. 1986 Mar;5(3):623–630. doi: 10.1002/j.1460-2075.1986.tb04256.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cole S. T., Brosch R., Parkhill J., Garnier T., Churcher C., Harris D., Gordon S. V., Eiglmeier K., Gas S., Barry C. E., 3rd Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature. 1998 Jun 11;393(6685):537–544. doi: 10.1038/31159. [DOI] [PubMed] [Google Scholar]
  10. Cole S. T., Eiglmeier K., Parkhill J., James K. D., Thomson N. R., Wheeler P. R., Honoré N., Garnier T., Churcher C., Harris D. Massive gene decay in the leprosy bacillus. Nature. 2001 Feb 22;409(6823):1007–1011. doi: 10.1038/35059006. [DOI] [PubMed] [Google Scholar]
  11. De Groote M. A., Ochsner U. A., Shiloh M. U., Nathan C., McCord J. M., Dinauer M. C., Libby S. J., Vazquez-Torres A., Xu Y., Fang F. C. Periplasmic superoxide dismutase protects Salmonella from products of phagocyte NADPH-oxidase and nitric oxide synthase. Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13997–14001. doi: 10.1073/pnas.94.25.13997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dussurget O., Stewart G., Neyrolles O., Pescher P., Young D., Marchal G. Role of Mycobacterium tuberculosis copper-zinc superoxide dismutase. Infect Immun. 2001 Jan;69(1):529–533. doi: 10.1128/IAI.69.1.529-533.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fang F. C., DeGroote M. A., Foster J. W., Bäumler A. J., Ochsner U., Testerman T., Bearson S., Giárd J. C., Xu Y., Campbell G. Virulent Salmonella typhimurium has two periplasmic Cu, Zn-superoxide dismutases. Proc Natl Acad Sci U S A. 1999 Jun 22;96(13):7502–7507. doi: 10.1073/pnas.96.13.7502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Farrant J. L., Sansone A., Canvin J. R., Pallen M. J., Langford P. R., Wallis T. S., Dougan G., Kroll J. S. Bacterial copper- and zinc-cofactored superoxide dismutase contributes to the pathogenesis of systemic salmonellosis. Mol Microbiol. 1997 Aug;25(4):785–796. doi: 10.1046/j.1365-2958.1997.5151877.x. [DOI] [PubMed] [Google Scholar]
  15. Figueroa-Bossi N., Bossi L. Inducible prophages contribute to Salmonella virulence in mice. Mol Microbiol. 1999 Jul;33(1):167–176. doi: 10.1046/j.1365-2958.1999.01461.x. [DOI] [PubMed] [Google Scholar]
  16. Figueroa-Bossi N., Uzzau S., Maloriol D., Bossi L. Variable assortment of prophages provides a transferable repertoire of pathogenic determinants in Salmonella. Mol Microbiol. 2001 Jan;39(2):260–271. doi: 10.1046/j.1365-2958.2001.02234.x. [DOI] [PubMed] [Google Scholar]
  17. Gilson E., Alloing G., Schmidt T., Claverys J. P., Dudler R., Hofnung M. Evidence for high affinity binding-protein dependent transport systems in gram-positive bacteria and in Mycoplasma. EMBO J. 1988 Dec 1;7(12):3971–3974. doi: 10.1002/j.1460-2075.1988.tb03284.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Harth G., Horwitz M. A. Export of recombinant Mycobacterium tuberculosis superoxide dismutase is dependent upon both information in the protein and mycobacterial export machinery. A model for studying export of leaderless proteins by pathogenic mycobacteria. J Biol Chem. 1999 Feb 12;274(7):4281–4292. doi: 10.1074/jbc.274.7.4281. [DOI] [PubMed] [Google Scholar]
  19. Hayashi S., Wu H. C. Lipoproteins in bacteria. J Bioenerg Biomembr. 1990 Jun;22(3):451–471. doi: 10.1007/BF00763177. [DOI] [PubMed] [Google Scholar]
  20. Hofmann K., Bucher P., Falquet L., Bairoch A. The PROSITE database, its status in 1999. Nucleic Acids Res. 1999 Jan 1;27(1):215–219. doi: 10.1093/nar/27.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hong Z., LoVerde P. T., Thakur A., Hammarskjöld M. L., Rekosh D. Schistosoma mansoni: a Cu/Zn superoxide dismutase is glycosylated when expressed in mammalian cells and localizes to a subtegumental region in adult schistosomes. Exp Parasitol. 1993 Mar;76(2):101–114. doi: 10.1006/expr.1993.1012. [DOI] [PubMed] [Google Scholar]
  22. Hoogenboom H. R., Griffiths A. D., Johnson K. S., Chiswell D. J., Hudson P., Winter G. Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains. Nucleic Acids Res. 1991 Aug 11;19(15):4133–4137. doi: 10.1093/nar/19.15.4133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jung J. U., Gutierrez C., Villarejo M. R. Sequence of an osmotically inducible lipoprotein gene. J Bacteriol. 1989 Jan;171(1):511–520. doi: 10.1128/jb.171.1.511-520.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Mariani F., Cappelli G., Riccardi G., Colizzi V. Mycobacterium tuberculosis H37Rv comparative gene-expression analysis in synthetic medium and human macrophage. Gene. 2000 Aug 8;253(2):281–291. doi: 10.1016/s0378-1119(00)00249-3. [DOI] [PubMed] [Google Scholar]
  26. Messing J., Gronenborn B., Müller-Hill B., Hans Hopschneider P. Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3642–3646. doi: 10.1073/pnas.74.9.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nielsen H., Engelbrecht J., Brunak S., von Heijne G. Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng. 1997 Jan;10(1):1–6. doi: 10.1093/protein/10.1.1. [DOI] [PubMed] [Google Scholar]
  28. Pesce A., Battistoni A., Stroppolo M. E., Polizio F., Nardini M., Kroll J. S., Langford P. R., O'Neill P., Sette M., Desideri A. Functional and crystallographic characterization of Salmonella typhimurium Cu,Zn superoxide dismutase coded by the sodCI virulence gene. J Mol Biol. 2000 Sep 15;302(2):465–478. doi: 10.1006/jmbi.2000.4074. [DOI] [PubMed] [Google Scholar]
  29. Pugsley A. P. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. doi: 10.1128/mr.57.1.50-108.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Raynaud C., Etienne G., Peyron P., Lanéelle M. A., Daffé M. Extracellular enzyme activities potentially involved in the pathogenicity of Mycobacterium tuberculosis. Microbiology. 1998 Feb;144(Pt 2):577–587. doi: 10.1099/00221287-144-2-577. [DOI] [PubMed] [Google Scholar]
  31. Riley L. W. Determinants of cell entry and intracellular survival of Mycobacterium tuberculosis. Trends Microbiol. 1995 Jan;3(1):27–31. doi: 10.1016/s0966-842x(00)88865-4. [DOI] [PubMed] [Google Scholar]
  32. San Mateo L. R., Toffer K. L., Orndorff P. E., Kawula T. H. Neutropenia restores virulence to an attenuated Cu,Zn superoxide dismutase-deficient Haemophilus ducreyi strain in the swine model of chancroid. Infect Immun. 1999 Oct;67(10):5345–5351. doi: 10.1128/iai.67.10.5345-5351.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Steinkühler C., Sapora O., Carrì M. T., Nagel W., Marcocci L., Ciriolo M. R., Weser U., Rotilio G. Increase of Cu,Zn-superoxide dismutase activity during differentiation of human K562 cells involves activation by copper of a constantly expressed copper-deficient protein. J Biol Chem. 1991 Dec 25;266(36):24580–24587. [PubMed] [Google Scholar]
  34. Takase I., Ishino F., Wachi M., Kamata H., Doi M., Asoh S., Matsuzawa H., Ohta T., Matsuhashi M. Genes encoding two lipoproteins in the leuS-dacA region of the Escherichia coli chromosome. J Bacteriol. 1987 Dec;169(12):5692–5699. doi: 10.1128/jb.169.12.5692-5699.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tatum F. M., Detilleux P. G., Sacks J. M., Halling S. M. Construction of Cu-Zn superoxide dismutase deletion mutants of Brucella abortus: analysis of survival in vitro in epithelial and phagocytic cells and in vivo in mice. Infect Immun. 1992 Jul;60(7):2863–2869. doi: 10.1128/iai.60.7.2863-2869.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wilks K. E., Dunn K. L., Farrant J. L., Reddin K. M., Gorringe A. R., Langford P. R., Kroll J. S. Periplasmic superoxide dismutase in meningococcal pathogenicity. Infect Immun. 1998 Jan;66(1):213–217. doi: 10.1128/iai.66.1.213-217.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wu C. H., Tsai-Wu J. J., Huang Y. T., Lin C. Y., Lioua G. G., Lee F. J. Identification and subcellular localization of a novel Cu,Zn superoxide dismutase of Mycobacterium tuberculosis. FEBS Lett. 1998 Nov 13;439(1-2):192–196. doi: 10.1016/s0014-5793(98)01373-8. [DOI] [PubMed] [Google Scholar]
  38. Zhang Y., Lathigra R., Garbe T., Catty D., Young D. Genetic analysis of superoxide dismutase, the 23 kilodalton antigen of Mycobacterium tuberculosis. Mol Microbiol. 1991 Feb;5(2):381–391. doi: 10.1111/j.1365-2958.1991.tb02120.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES