1887

Abstract

Summary: Several species of the opportunistic fungal pathogen produce an extracellular aspartyl proteinase that may assist the organism to invade and colonize host tissues, evade the host immune response and assimilate nitrogen from proteinaceous sources. Although addition of exogenous proteins, such as bovine serum albumin (BSA), to cultures of is known to elicit proteinase production, the precise molecular mechanisms controlling regulation of proteinase induction are unknown. We have examined the ability of a variety of macromolecules to induce proteinase production using a chemically-defined nitrogen-limited growth medium and a rapid, sensitive microtitre fluorescent assay for proteinase activity in culture supernatants. BSA and the extracellular matrix protein collagen induced proteinase production. Homopolymers of both poly-L- and poly-D-glutamate also induced proteinase activity, whereas polyglycine, heparin sulphate and dextran sulphate did not. Thus, molecular recognition of proteinase-inducing stimuli is not highly stereospecific, but apparently requires both main- and side-chain interactions. Peptides 8 or more residues in length generally induced proteinase production while most shorter peptides did not. These data reveal that internalization of small peptides with less than 7 residues by peptide transport was not the inducing signal for proteinase production, since dipeptide and oligopeptide permeases do not efficiently transport peptides of more than 6-7 residues. In addition a tight-binding synthetic inhibitor of Candida proteinase ( = 0.17 nM) prevented growth of on BSA as a sole nitrogen source by blocking protein degradation. Immunodetection of proteinase in these culture supernatants suggests that fully intact proteins, in addition to peptide fragments of sufficient size, are capable of inducing proteinase production. A model involving stimulation of a plasma membrane signal transduction event by extracellular protein and/or polypeptide ligands of more than seven residues is compatible with these data.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-7-1643
1993-07-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/7/mic-139-7-1643.html?itemId=/content/journal/micro/10.1099/00221287-139-7-1643&mimeType=html&fmt=ahah

References

  1. Ausubel F.M., Brent R., Kingston R.E., Moore D.D., Seidman J.G., Smith J.A., Struhl K. 1990; Saccharomyces cerevisiae.. Current Protocols in Molecular Biology 2:13.1.3
    [Google Scholar]
  2. Banerjee A., Ganesan K., Datta A. 1991; Induction of secretory acid proteinase in Candida albicans.. Journal of General Microbiology 137:2455–2461
    [Google Scholar]
  3. Basrai M.M., Naider F., Becker J.M. 1990; Internalization of lucifer yellow in Candida albicans by fluid phase endocytosis.. Journal of General Microbiology 136:1059–1065
    [Google Scholar]
  4. Borg M., Ruchel R. 1988; Expression of extracellular acid proteinase by proteolytic Candida during experimental infection of oral mucosa.. Infection and Immunity 56:626–631
    [Google Scholar]
  5. Borg M., Ruchel R. 1990; Demonstration of fungal proteinase during phagocytosis of Candida albicans and Candida tropicalis.. Journal of Medical and Veterinary Mycology 28:3–14
    [Google Scholar]
  6. Capobianco J.O., Lerner G.G., Goldman R.C. 1992; Application of a fluorogenic substrate in the assay of proteolytic activity and in the discovery of a potent inhibitor of Candida albicansaspartic proteinase.. Analytical Biochemistry 203:96–102
    [Google Scholar]
  7. Crandall M., Edwards J.E. 1987; Segregation of proteinasenegative mutants from heterozygous Candida albicans.. Journal of General Microbiology 133:2817–2824
    [Google Scholar]
  8. Cutler J.E. 1991; Putative virulence factors of Candida albicans. . Annual Review of Microbiology 45:187–218
    [Google Scholar]
  9. De Bernardis F., Agatensi L., Ross I.K., Emerson G.W., Lorenzini R., Sullivan P.A., Cassone A. 1990; Evidence for a role for secreted aspartyl proteinase of Candida albicans in vulvovaginal candidiasis.. Journal of Infectious Diseases 161:1276–1283
    [Google Scholar]
  10. Douglas L.J. 1988; Candida proteinases and candidosis.. CRC Critical Reviews in Microbiology 8:121–129
    [Google Scholar]
  11. Ganesan K., Banerjee A., Datta A. 1991; Molecular cloning of the secretory acid proteinase gene from Candida albicans and its use as a species-specific probe.. Infection and Immunity 59:2972–2977
    [Google Scholar]
  12. Ghannoum M., Abu Elteen K. 1986; Correlative relationship between proteinase production, adherence and pathogenicity of various strains of Candida albicans.. Journal of General Microbiology 129:431–438
    [Google Scholar]
  13. Hattori M., Yoshiura K., Negi M., Ogawa H. 1984; Keratinolytic proteinase produced by Candida albicans.. Sabouraudia 22:175–183
    [Google Scholar]
  14. Homma M., Kanbe T., Chibana H., Tanaka K. 1992; Detection of intracellular forms of secretory aspartic proteinase in Candida albicans.. Journal of General Microbiology 138:627–633
    [Google Scholar]
  15. Hube B., Turver C.J., Odds F.C., Eiffert H., Boulnois G.J., Kochel H., Ruchel R. 1991; Sequence of the Candida albicans gene encoding the secretory aspartate proteinase.. Journal of Medical and Veterinary Mycology 29:129–132
    [Google Scholar]
  16. Island M.D., Naider F., Becker J.M. 1987; Regulation of dipeptide transport in Saccharomyces cerevisiae by micromolar amino acid concentrations.. Journal of Bacteriology 169:2132–2136
    [Google Scholar]
  17. Kaminishi H., Hagihara Y., Hayashi S., Cho T. 1986; Isolation and characteristics of collagenolytic enzyme produced by Candida albicans.. Infection and Immunity 53:312–316
    [Google Scholar]
  18. Kaminishi H., Hagihara Y., Tanaka M., Cho T. 1988; Degradation of bovine achilles tendon collagen by Candida albicansproteinase.. Journal of Medical and Veterinary Mycology 26:315–318
    [Google Scholar]
  19. Kaminishi H., Tanaka M., Cho T., Maeda H., Hagihara Y. 1990; Activation of the plasma kallikrein-kinin system by Candida albicans proteinase.. Infection and Immunity 58:2139–2143
    [Google Scholar]
  20. Kondoh Y., Shimizu K., Tanaka K. 1987; Proteinase production and pathogenicity of Candida albicans. II. Virulence for mice of C. albicans strains of different proteinase activity.. Microbiology and Immunology 31:1061–1069
    [Google Scholar]
  21. Kwon-Chung K.J., Lehman D., Good C., Magee P.T. 1985; Genetic evidence for role of extracellular proteinase in virulence of Candida albicans.. Infection and Immunity 49:571–575
    [Google Scholar]
  22. Mccarthy P.J., Nisbet L.J., Boehm J.C., Kingsbury W.D. 1985; Multiplicity of peptide permeases in Candida albicans:evidence from novel chromogenic peptides.. Journal of Bacteriology 162:1024–1029
    [Google Scholar]
  23. Macdonald F. 1984; Secretion of inducible proteinase by pathogenic Candida species.. Journal of Medical and Veterinary Mycology 22:79–82
    [Google Scholar]
  24. Macdonald F., Odds F.C. 1980; Inducible proteinase of Candida albicans in diagnostic serology and the pathogenesis of systemic candidosis.. Journal of Medical Microbiology 13:423–435
    [Google Scholar]
  25. Macdonald F., Odds F.C. 1983; Virulence for mice of a proteinase-secreting strain of Candida albicans and a proteinase-deficient mutant.. Journal of General Microbiology 129:431–438
    [Google Scholar]
  26. Milewski S., Andruskiewicz R., Borowski E. 1988; Substrate specificity of peptide transport in Candida albicans.. FEMS Microbiology Letters 50:73–78
    [Google Scholar]
  27. Morrow B., Srikantha T., Soll D.R. 1992; Transcription of the gene for a pepsinogen, PEP1, is regulated by white-opaque switching in Candida albicans.. Molecular and Cellular Biology 12:2997–3005
    [Google Scholar]
  28. Naider F., Becker J.M. 1988; Peptide transport in Candida albicans: implications for the development of antifungal agents.. Current Topics in Medical Microbiology 2:170–198
    [Google Scholar]
  29. Odds F.C. 1985; Candida albicans proteinase as a virulence factor in the pathogenesis of Candida infections.. Zentralblatt für Bakteriologie, Mikrobiologie und Hygiene (Abteilung I) 260:539–542
    [Google Scholar]
  30. Payne J.W. 1986; Drug delivery systems: optimising the structure of peptide carriers for synthetic antimicrobial drugs.. Drugs Experimental Clinical Research 12:585–594
    [Google Scholar]
  31. Payne J.W., Shallow D.A. 1985; Studies on drug targeting in the pathogenic fungus Candida albicans: peptide transport mutants resistant to polyoxins, nikkomycins and bacilysin.. FEMS Microbiology Letters 28:55–65
    [Google Scholar]
  32. Payne J.W., Barrett-Bee K.J., Shallow D.A. 1991; Peptide substrates rapidly modulate expression of dipeptide and oligopeptide permease in Candida albicans.. FEMS Microbiology Letters 79:15–20
    [Google Scholar]
  33. Raths S.K., Naider F., Becker J.M. 1988; Peptide analogs compete with the binding of alpha-factor to its receptor in Saccharomyces cerevisiae.. Journal of Biological Chemistry 263:17333–17341
    [Google Scholar]
  34. Ray T.L., Payne C.D. 1990; Comparative production and rapid purification of Candida acid proteinase from protein-supplemented cultures.. Infection and Immunity 58:508–514
    [Google Scholar]
  35. Remold H., Fasold H., Staib F. 1968; Purification and characterization of a proteolytic enzyme from Candida albicans.. Biochimica et Biophysica Acta 167:399–406
    [Google Scholar]
  36. Ross I.K., Debernardis F., Emerson G.W., Cassone A., Sullivan P.A. 1990; The secreted aspartate proteinase of Candida albicans: physiology of secretion and virulence of a proteinase- deficient mutant.. Journal of General Microbiology 136:687–694
    [Google Scholar]
  37. Ruchel R. 1981; Properties of a purified proteinase from the yeast Candida albicans.. Biochimica et Biophysica Acta 659:99–113
    [Google Scholar]
  38. Ruchel R. 1983; On the role of proteinases from Candida albicans in the pathogenesis of acronecrosis.. Zentralblatt für Bakteriologie, Mikrobiologie und Hygiene (Abteilung I) 255:524–537
    [Google Scholar]
  39. Ruchel R. 1986; Cleavage of immunoglobulins by pathogenic yeasts of the genus Candida.. Microbiological Sciences 3:316–319
    [Google Scholar]
  40. Ruchel R., Uhlemann K., Boning B. 1983; Secretion of acid proteinases by different species of the genus Candida.. Zentralblatt für Bakteriologie, Mikrobiologie und Hygiene (Abteilung I) 255:537–548
    [Google Scholar]
  41. Ruchel R., Boning B., Borg M. 1986; Characterization of a secretory proteinase of Candida parapsilosis and evidence for the absence of the enzyme during infection in vitro.. Infection and Immunity 53:411–419
    [Google Scholar]
  42. Ruchel R., Zimmermann F., Boning-Stutzer B., Helmchen U. 1991; Candidiasis visualized by proteinase-directed immuno-fluorescence.. Virchows Archiv A, Pathological Anatomy and Histopathology 419:199–202
    [Google Scholar]
  43. Sadhu C., Hoekstra D., Mceachern M.J., Reed S.I., Hicks J.B. 1992; A G-protein ɑ subunit from asexual Candida albicansfunctions in the mating signal transduction pathway of Saccharomyces cerevisiae and is regulated by the al-ɑ2 repressor.. Molecular and Cellular Biology 12:1977–1985
    [Google Scholar]
  44. Shallow D.A., Barrett-Bee K.J., Payne J.W. 1991; Evaluation of the dipeptide and oligopeptide permeases of Candida albicans as uptake routes for synthetic anticandidal agents.. FEMS Microbiology Letteganeshrs 79:9–14
    [Google Scholar]
  45. Staib F. 1965; Serum-protein as nitrogen source for yeast-like fungi.. Journal of Medical and Veterinary Mycology 4:187–193
    [Google Scholar]
  46. Togni G., Sanglard D., Falchetto R., Monod M. 1991; Isolation and nucleotide sequence of the extracellular acid protease gene (ACP) from the yeast Candida tropicalis.. FEBS Letters 286:181–185
    [Google Scholar]
  47. Wright R.J., Carne A., Hieber A.D., Lamont I.L., Emerson G.W., Sullivan P.A. 1992; A second gene for a secreted aspartate proteinase in Candida albicans.. Journal of Bacteriology 174:7848–7853
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-7-1643
Loading
/content/journal/micro/10.1099/00221287-139-7-1643
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