Identification of wall-specific antigens synthesized during germ tube formation by Candida albicans.

摘要: Walls of the two cellular forms (blastoconidia and mycelia) Candida albicans ATCC 26555 were obtained from cells metabolically labeled (6-h pulse) with 14C-protein hydrolysate [3H]threonine. purified by thorough washings buffered sodium dodecyl sulfate solutions digested Zymolyase 20T. The enzymatic treatment released four major high-molecular-weight mannoproteins (HMWM), apparent molecular masses 650, 500, 340, 200 kilodaltons (HMWM-650, HMWM-500, HMWM-340, HMWM-200, respectively), yeast cells, whereas high-molecular-mass (HMWM-260 HMWM-180) solubilized mycelial cells. Some additional minor low-molecular-weight species also detected in digests walls both types cell. Single dual pulse-chase experiments indicated that HMWM-260 HMWM-180 reflect de novo synthesis new proteins specific for mycelia do not represent a topological rearrangement blastoconidium wall components. Monoclonal antibodies raised against (quantitatively predominant component walls), polyclonal rabbit or cell walls. Anti-mycelial adsorbed to whole killed blastoconidia remove common antigens. Titration enzyme-linked immunosorbent assay revealed monoclonal could recognize an epitope protein moiety mannoprotein. Immunoblotting immunofluorescence techniques using these confirmed are components envelope Images