Identification of a 58-kilodalton cell surface fibrinogen-binding mannoprotein from Candida albicans.
Treatment of both yeast (blastoconidia) and hyphal (blastoconidia with germ tubes) cells of Candida albicans with beta-mercaptoethanol (beta ME) releases a complex array of cell wall-bound proteins and glycoproteins. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblotting with fibrinogen-anti-fibrinogen antibody allowed the identification of a 58-kDa mannoprotein (mp58) in both extracts which specifically interacts with human fibrinogen. Treatment of intact cells with low concentrations of beta-glucanase (Zymolyase 20T) for short periods or with beta ME abolished or significantly reduced binding of fibrinogen. A rabbit polyclonal antiserum was raised against the purified mp58 species released by beta ME from germinated blastoconidia (PAb anti-mp58). By Western blotting, the antiserum cross-reacted with the homologous 58-kDa fibrinogen-binding mannoprotein present in beta ME extracts from blastoconidia, and by indirect immunofluorescence, the antiserum labelled both yeast cells and hyphae, yet reactivity was found primarily on the cell surface of filamentous forms. Immunostaining of human infected tissue sections with PAb anti-mp58 showed that the mp58 species is also expressed in vivo; in this case, the species is in the forms of both yeast and hyphal elements similarly labelled by the antiserum. Purified immunoglobulin G fraction from the antiserum did not alter the binding of fibrinogen as determined by a modified enzyme-linked immunosorbent assay and Western blotting. The N- and O-glycosidically linked carbohydrates represent 18 to 20% and 3 to 4%, respectively, of the molecular mass of the mp58. O-linked sugar residues may be involved in the interaction of the molecule with fibrinogen.
Casanova
M
M
Sección Departamental de Microbiología, Facultad de Farmacia, Universitat de Valencia, Spain.
Lopez-Ribot
J L
JL
Monteagudo
C
C
Llombart-Bosch
A
A
Sentandreu
R
R
Martinez
J P
JP
D000818:Animals; D002176:Candida albicans / Q000737:chemistry*; D005340:Fibrinogen / Q000378:metabolism*; D005656:Fungal Proteins / Q000302:isolation & purification*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D008623:Mercaptoethanol / Q000494:pharmacology; D011817:Rabbits
D005656:Fungal Proteins; D007106:Immune Sera; D008562:Membrane Glycoproteins; C025363:mannoproteins; D008623:Mercaptoethanol; D005340:Fibrinogen