1887

Abstract

isolates of genomovar III are highly transmissible amongst patients with cystic fibrosis (CF) and express a 97 kDa putative haem-binding protein (HBP) [ Smalley, J. W., Charalabous, P., Birss, A. J. & Hart, C. A. (2001) . , 509–514]. An investigation of the interactions of iron(III) protoporphyrin IX with epidemic and non-epidemic strains of to determine the role of the above protein in haem acquisition and binding is reported herein. Spectrophotometric titrations of cell suspensions of genomovar IIIa strains BC7 and C5424 with iron(III) protoporphyrin IX, at pH 7·0, resulted in the depletion of Fe(III)PPIX.OH monomers and formation of the -oxo oligomeric species, [Fe(III)PPIX]O. Difference spectroscopy indicated a continuous conversion of the monomeric iron(III) protoporphyrin IX into -oxo oligomers. Incubations with Fe(III)PPIX.OH monomers at pH 6·5 also showed that cells could shift the equilibrium to generate the -oxo oligomeric form. Genomovar I strains ATCC 25416 and LMG 17997 were unable to mediate this conversion. SDS-PAGE of genomovar IIIa strains exposed to Fe(III)PPIX.OH at pH 6·5 followed by tetramethylbenzidine/HO staining revealed, in addition to the 97 kDa HBP, two proteins of 77 and 149 kDa located in the outer membrane which bound Fe(III)PPIX.OH monomers. These proteins were absent from the genomovar I strains. Genomovar IIIa strains BC7 and C5424 showed increased cellular binding of [Fe(III)PPIX]O, and as a consequence, displayed increased catalase activities compared to cells of the genomovar I isolates. It is concluded that, in addition to the putative 97 kDa HBP, genomovar IIIa strains express two outer-membrane proteins which function to bind and convert Fe(III)PPIX.OH monomers into the -oxo oligomeric form, [Fe(III)PPIX]O. The ability to perform this conversion at both neutral and slightly acidic pHs may enable epidemic strains to withstand attack from neutrophil-derived HO in the inflamed CF lung.

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2003-04-01
2020-01-21
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References

  1. Barnard M. L., Muller-Eberhard U., Turrens J. F.. 1993; Protective role of hemopexin on heme-dependent lung oxidative stress. Biochem Biophys Res Commun192:82–87
    [Google Scholar]
  2. Beers R. F., Sizer I. W.. 1952; A spectrophotometric assay for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem196:133–140
    [Google Scholar]
  3. Bodem C. R., Lampton L. M., Miller D. P., Tarka E. F., Everett E. D.. 1983; Endobronchial pH relevance to aminoglycoside activity in gram-negative bacillary pneumonia. Am Rev Respir Dis127:39–41
    [Google Scholar]
  4. Brown S. B., Dean T. C., Jones P.. 1970; Catalytic activity of iron(III)-centred catalysts. Role of dimerization in the catalytic actions of ferrihaems. Biochem J117:741–744
    [Google Scholar]
  5. Chester B.. 1979; Semiquantitative catalase test as an aid in identification of oxidative and nonsaccharolytic Gram-negative bacteria. J Clin Microbiol10:525–528
    [Google Scholar]
  6. Gessner A. R., Mortensen J. E.. 1990; Pathogenic factors of Pseudomonas cepacia isolates from patients with cystic fibrosis. J Med Microbiol33:115–120
    [Google Scholar]
  7. Grinberg L. N., O'Brien P. J., Hrkal Z.. 1999; The effects of heme-binding proteins on the peroxidative and catalytic activities of hemin. Free Rad Biol Med26:214–219
    [Google Scholar]
  8. Gutteridge J. M. C., Smith A.. 1988; Antioxidant protection by haemopexin of haem-stimulated lipid peroxidation. Biochem J256:861–865
    [Google Scholar]
  9. Hart C. A., Winstanley C.. 2002; Persistent and aggressive bacteria in the lungs of cystic fibrosis children. Br Med Bull61:81–96
    [Google Scholar]
  10. Jayaraman S., Joo N. S., Reitz B., Wine J. J., Verkman A. S.. 2001; Submucosal gland secretions in airways from cystic fibrosis patients have normal [Na+] and pH but elevated viscosity. Proc Nat Acad Sci U S A98:8119–8123
    [Google Scholar]
  11. Jones P., Robson T., Brown S. B.. 1973; The catalase activity of ferrihaems. Biochem J135:353–359
    [Google Scholar]
  12. Lee B. C.. 1992; Isolation of haemin-binding proteins of Neisseria gonorrhoeae . J Med Microbiol36:121–127
    [Google Scholar]
  13. Lefebre M. D., Valvano M. A.. 2001; In vitro resistance of Burkholderia cepacia complex isolates to reactive oxygen species in relation to catalase and superoxide dismutase production. Microbiology147:97–109
    [Google Scholar]
  14. Liem H. H., Tavassoli M., Muller-Eberhard U.. 1975; Cellular and subcellular localization of heme and hemopexin in the rabbit. Acta Haematol53:219–225
    [Google Scholar]
  15. Mahenthiralingam E., Coenye T., Chung J. W., Speert D. P., Govan J. R. W., Taylor P., Vandamme P.. 2000; Diagnostically and experimentally useful panel of strains from the Burkholderia cepacia complex. J Clin Microbiol38:910–913
    [Google Scholar]
  16. Mahenthiralingam E., Vandamme P., Campbell M. E.. 7 other authors 2001; Infection with Burkholderia cepacia complex genomovars in patients with cystic fibrosis: virulent transmissible strains of genomovar III can replace Burkholderia multivorans . Clin Infect Dis33:1469–1475
    [Google Scholar]
  17. Mazoy R., Lemos M. L.. 1996; Identification of heme-binding proteins in the cell membranes of Vibrio anguillarum . FEMS Microbiol Lett135:265–270
    [Google Scholar]
  18. Miller J. R., Taies J. A., Silver J.. 1987; Mossbauer and spectroscopic studies on substituted tetraphenylporphyrinato iron (III) complexes in aqueous solutions and the formation of the μ -oxo-bridged species. Inorg Chim Acta138:205–214
    [Google Scholar]
  19. Silver J., Lukas B.. 1983; Mössbauer studies on protoporphyrin IX iron(III) solutions. Inorg Chim Acta78:219–224
    [Google Scholar]
  20. Smalley J. W., Birss A. J., McKee A. S., Marsh P. D.. 1993; Haemin-binding proteins in Porphyromonas gingivalis W50 grown in the chemostat under haemin-limitation. J Gen Microbiol139:2145–2150
    [Google Scholar]
  21. Smalley J. W., Silver J., Marsh P. J., Birss A. J.. 1998; The periodontopathogen Porphyromonas gingivalis binds iron protoporphyrin IX in the μ -oxo dimeric form: an oxidative buffer and possible pathogenic mechanism. Biochem J331:681–677
    [Google Scholar]
  22. Smalley J. W., Birss A. J., Silver J.. 2000; The periodontal pathogen Porphyromonas gingivalis harnesses the chemistry of the μ -oxo bishaem of iron protoporphyrin IX to protect against hydrogen peroxide. FEMS Microbiol Lett183:159–164
    [Google Scholar]
  23. Smalley J. W., Charalabous P., Birss A. J., Hart C. A.. 2001; Detection of heme-binding proteins in epidemic strains of Burkolderia cepacia . Clin Diag Lab Immunol8:509–514
    [Google Scholar]
  24. Smalley J. W., Birss A. J., Withnall R., Silver J.. 2002; Interactions of Porphyromonas gingivalis with oxyhaemoglobin and deoxyhaemoglobin. Biochem J362:239–245
    [Google Scholar]
  25. Stugard C. E., Daskaleros P. A., Pain S. M.. 1989; A 101-kilodalton heme-binding protein associated with congo red binding and virulence of Shigella flexneri and enteroinvasive Escherichia coli strains. Infect Immun57:3534–3539
    [Google Scholar]
  26. Wandersman C., Stojiljkovic I.. 2000; Bacterial heme sources: the role of heme, hemoprotein receptors and hemophores. Curr Opin Microbiol3:215–220
    [Google Scholar]
  27. White W. I.. 1978; In The Porphyrins vol. 7 p303 Edited by Dolphin D.. London: Academic Press;
    [Google Scholar]
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