title: Partial biochemical characterization of cell surface hydrophobicity and hydrophilicity of Candida albicans.
abstract: Hydrophobic yeast cells of Candida albicans are more virulent than hydrophilic yeast cells in mice. Results of experiments performed in vitro suggest that surface hydrophobicity contributes to virulence in multiple ways. Before definitive studies in vivo concerning the contribution of fungal surface hydrophobicity to pathogenesis can be performed, biochemical, physiological, and immunochemical characterization of the macromolecules responsible for surface hydrophobicity must be accomplished. This report describes our initial progress toward this goal. When hydrophobic and hydrophilic yeast cells of C. albicans were exposed to various enzymes, only proteases caused any change in surface hydrophobicity. Hydrophobic cell surfaces were sensitive to trypsin, chymotrypsin, pronase E, and pepsin. This indicates that surface hydrophobicity is due to protein. Papain, however, had no significant effect. The hydrophobicity of hydrophilic cells was altered only by papain. The proteins responsible for surface hydrophobicity could be removed by exposure to lyticase, a beta 1-3 glucanase, for 30 to 60 min. When 60-min lyticase digests of hydrophobic and hydrophilic cell walls were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a 12.5% resolving gel, each protein population contained a single unique protein that was not evident in the other protein population. However, when the cell wall surface proteins of hydrophobic and hydrophilic cells were first labeled with 125I and then removed by lyticase and analyzed by SDS-PAGE, at least four low-molecular-mass (less than 65 kilodaltons) proteins associated with hydrophobic cells were either absent or much less abundant in the hydrophilic cell digests. This result was seen for both C. albicans strains that we tested. When late-exponential-phase hydrophilic cells were treated with tunicamycin, high levels of surface hydrophobicity were obtained by stationary phase. These results indicate that the surface hydrophobicity of C. albicans reflects changes in external surface protein exposure and that protein mannosylation may influence exposure of hydrophobic surface proteins.
authors.affiliation: Division of Biological Sciences, University of Montana, Missoula 59812.
authors.forename: K C
authors.initials: KC
authors.lastname: Hazen
authors.forename: J G
authors.initials: JG
authors.lastname: Lay
authors.forename: B W
authors.initials: BW
authors.lastname: Hazen
authors.forename: R C
authors.initials: RC
authors.lastname: Fu
authors.forename: S
authors.initials: S
authors.lastname: Murthy
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D002176:Candida albicans / Q000032:analysis* / Q000187:drug effects; D002473:Cell Wall / Q000737:chemistry* / Q000187:drug effects; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000494:pharmacology; D007456:Iodine Isotopes; D009097:Multienzyme Complexes / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000494:pharmacology; D012995:Solubility; D013499:Surface Properties; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D001425:Bacterial Outer Membrane Proteins; D007456:Iodine Isotopes; D009097:Multienzyme Complexes; C025310:lyticase; D014415:Tunicamycin; D004693:Glucan Endo-1,3-beta-D-Glucosidase; D010447:Peptide Hydrolases; D004229:Dithiothreitol