1887

Abstract

In this study, extracellular glutathione-dependent ferric reductase (GSH-FeR) activities in different dimorphic zoopathogenic fungal species were characterized. Supernatants from , , and strains grown in their yeast form were able to reduce iron enzymically with glutathione as a cofactor. Some variations in the level of reduction were noted amongst the strains. This activity was stable in acidic, neutral and slightly alkaline environments and was inhibited when trivalent aluminium and gallium ions were present. Using zymography, single bands of GSH-FeRs with apparent molecular masses varying from 430 to 460 kDa were identified in all strains. The same molecular mass range was determined by size exclusion chromatography. These data demonstrate that dimorphic zoopathogenic fungi produce and secrete a family of similar GSH-FeRs that may be involved in the acquisition and utilization of iron. Siderophore production by these and other fungi has sometimes been considered to provide a full explanation of iron acquisition in these organisms. Our work reveals an additional common mechanism that may be biologically and pathogenically important. Furthermore, while some characteristics of these enzymes such as extracellular location, cofactor utilization and large size are not individually unique, when considered together and shared across a range of fungi, they represent an important novel physiological feature.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.27918-0
2005-07-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/151/7/mic1512233.html?itemId=/content/journal/micro/10.1099/mic.0.27918-0&mimeType=html&fmt=ahah

References

  1. Adams T. J., Vartivarian S., Cowart R. E. 1990; Iron acquisition systems of Listeria monocytogenes . Infect Immun 58:2715–2718
    [Google Scholar]
  2. Chen S. C. A., Wright L. C., Santangelo R. T., Muller M., Moran V. R., Kuchel P. W., Sorrell T. C. 1997; Identification of extracellular phospholipase B, lysophospholipase, and acyltransferase produced by Cryptococcus neoformans . Infect Immun 65:405–411
    [Google Scholar]
  3. Chitambar C. R., Matthaeus W. G., Antholine W. E., Graff K., O'Brien W. J. 1988; Inhibition of leukemic HL60 cell growth by transferrin-gallium: effects on ribonucleotide reductase and demonstration of drug synergy with hydroxyurea. Blood 72:1930–1936
    [Google Scholar]
  4. Cowart R. E. 2002; Reduction of iron by extracellular iron reductases: implications for microbial iron acquisition. Arch Biochem Biophys 400:273–281 [CrossRef]
    [Google Scholar]
  5. de Luca N. G., Wood P. M. 2000; Iron uptake by fungi: contrasted mechanisms with internal or external reduction. Adv Microb Physiol 43:39–74
    [Google Scholar]
  6. Deneer H. G., Boychuk I. 1993; Reduction of iron by Listeria monocytogenes and other species of Listeria . Can J Microbiol 39:480–485 [CrossRef]
    [Google Scholar]
  7. Eissenberg L. G., West J. L., Woods J. P., Goldman W. E. 1991; Infection of P388D1 macrophages and respiratory epithelial cells by Histoplasma capsulatum : selection of avirulent variants and their potential role in persistent histoplasmosis. Infect Immun 59:1639–1646
    [Google Scholar]
  8. Foster L. A. 2002; Utilization and cell-surface binding of hemin by Histoplasma capsulatum . Can J Microbiol 48:437–442 [CrossRef]
    [Google Scholar]
  9. Homuth M., Valentin-Weigand P., Rohde M., Gerlach G.-F. 1998; Identification and characterization of a novel extracellular ferric reductase from Mycobacterium paratuberculosis . Infect Immun 66:710–716
    [Google Scholar]
  10. Howard D. H. 1999; Acquisition, transport, and storage of iron by pathogenic fungi. Clin Microbiol Rev 12:394–404
    [Google Scholar]
  11. Howard D. H., Rafie R., Tiwari A., Faull K. F. 2000; Hydroxyamate siderophores of Histoplasma capsulatum . Infect Immun 68:2338–2343 [CrossRef]
    [Google Scholar]
  12. Kaufmann F., Lovley D. R. 2001; Isolation and characterization of a soluble NADPH-dependent Fe(III) reductase from Geobacter sulfurreducens . J Bacteriol 183:4468–4476 [CrossRef]
    [Google Scholar]
  13. Kosman D. J. 2003; Molecular mechanisms of iron uptake in fungi. Mol Microbiol 47:1185–1197 [CrossRef]
    [Google Scholar]
  14. Laemmli U. K. 1970; Cleaving of the structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
    [Google Scholar]
  15. Lancashire R. J. 2002; Chemistry of iron. www.chem.uwimona.edu.jm:1104/courses/iron.html
  16. Mazoch J., Tesařik R., Sedláček V., Kučera I., Turánek J. 2004; Isolation and biochemical characterization of two soluble iron(III) reductases from Paracoccus denitrificans . Eur J Biochem 271:553–562 [CrossRef]
    [Google Scholar]
  17. Mazoy R., Lopez E. M., Fouz B., Amaro C., Lemos M. L. 1999; Ferric-reductase activities in Vibrio vulnificus biotypes 1 and 2. FEMS Microbiol Lett 172:205–211 [CrossRef]
    [Google Scholar]
  18. Morrissey J. A., Williams P. H., Cashmore A. M. 1996; Candida albicans has a cell-associated ferric-reductase activity, which is regulated in response to levels of iron and copper. Microbiology 142:485–492 [CrossRef]
    [Google Scholar]
  19. Newman S. L., Gootee L., Brunner G., Deepe G. S. Jr 1994; Chloroquine induces human macrophage killing of Histoplasma capsulatum by limiting the availability of intracellular iron and is therapeutic in a murine model of histoplasmosis. J Clin Invest 93:1422–1429 [CrossRef]
    [Google Scholar]
  20. Nyhus K. J., Wilborn A. T., Jacobson E. S. 1997; Ferric iron reduction by Cryptococcus neoformans . Infect Immun 65:434–438
    [Google Scholar]
  21. Olakanmi O., Britigan B. E., Schlesinger L. S. 2000; Gallium disrupts iron metabolism of mycobacteria residing within human macrophages. Infect Immun 68:5619–5627 [CrossRef]
    [Google Scholar]
  22. Poch M. T., Johnson W. 1993; Ferric-reductases of Legionella pneumophila . BioMetals 6:107–114
    [Google Scholar]
  23. Ramanan N., Wang Y. 2000; A high-affinity iron permease essential for Candida albicans virulence. Science 288:1062–1064 [CrossRef]
    [Google Scholar]
  24. Schröder I., Johnson E., de Vries S. 2003; Microbial ferric iron reductases. FEMS Microbiol Rev 27:427–447 [CrossRef]
    [Google Scholar]
  25. Timmerman M. M., Woods J. P. 1999; Ferric reduction is a potential iron acquisition mechanism for Histoplasma capsulatum . Infect Immun 67:6403–6408
    [Google Scholar]
  26. Timmerman M. M., Woods J. P. 2001; Potential role for extracellular glutathione-dependent ferric reductase in utilization of environmental and host ferric compounds by Histoplasma capsulatum . Infect Immun 69:7671–7678 [CrossRef]
    [Google Scholar]
  27. Tsuboi R., Sanada T., Takamori K., Ogawa H. 1987; Isolation and properties of extracellular proteinases from Sporothrix schenckii . J Bacteriol 169:4104–4109
    [Google Scholar]
  28. Vadas A., Monbouquette H. G., Johnson E., Schröder I. 1999; Identification and characterization of a novel ferric reductase from the hyperthermophilic archeon Archaeoglobus fulgidus . J Biol Chem 274:36715–36721 [CrossRef]
    [Google Scholar]
  29. Weinberg E. D. 1999; The role of iron in protozoan and fungal infectious diseases. J Eukaryot Microbiol 46:231–238 [CrossRef]
    [Google Scholar]
  30. Woods J. P. 2003; Knocking on the right door and making a comfortable home: Histoplasma capsulatum intracellular pathogenesis. Curr Opin Microbiol 6:327–331 [CrossRef]
    [Google Scholar]
  31. Woods J. P., Heinecke E. L., Goldman W. E. 1998; Electrotransformation and expression of bacterial genes encoding hygromycin phosphotransferase and β -galactosidase in the pathogenic fungus Histoplasma capsulatum . Infect Immun 66:1697–1707
    [Google Scholar]
  32. Wulfsberg G. 2000 Table C. In Inorganic Chemistry , inside back cover Sausalito: University Science Books;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.27918-0
Loading
/content/journal/micro/10.1099/mic.0.27918-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error