title: Identification of wall-specific antigens synthesized during germ tube formation by Candida albicans.
abstract: Walls of the two cellular forms (blastoconidia and mycelia) of Candida albicans ATCC 26555 were obtained from cells metabolically labeled (6-h pulse) with 14C-protein hydrolysate and [3H]threonine. Walls were purified by thorough washings with buffered and sodium dodecyl sulfate solutions and digested with Zymolyase 20T. The enzymatic treatment released four major high-molecular-weight mannoproteins (HMWM), with apparent molecular masses of 650, 500, 340, and 200 kilodaltons (HMWM-650, HMWM-500, HMWM-340, and HMWM-200, respectively), from yeast cells, whereas two high-molecular-mass mannoproteins (HMWM-260 and HMWM-180) were solubilized from mycelial cells. Some additional minor low-molecular-weight species were also detected in the enzymatic digests of walls from both types of cell. Single and dual pulse-chase experiments indicated that the HMWM-260 and HMWM-180 species reflect de novo synthesis of new proteins specific for the mycelia and do not represent a topological rearrangement of blastoconidium wall components. Monoclonal antibodies were raised against the HMWM-260 species (quantitatively the predominant component in the mycelial walls), and polyclonal rabbit antibodies were obtained against yeast or mycelial cell walls. Anti-mycelial cell wall polyclonal antibodies were adsorbed to whole killed blastoconidia to remove antibodies against common blastoconidium and mycelial wall antigens. Titration by enzyme-linked immunosorbent assay revealed that the monoclonal antibodies could recognize an epitope of the protein moiety of the HMWM-260 mannoprotein. Immunoblotting and immunofluorescence techniques using these monoclonal and polyclonal antibodies confirmed that the HMWM-260 and HMWM-180 species are specific components of the envelope of the mycelial cell walls.
authors: Departamento de Micrbiología, Facultad de Farmacia, Valencia, Spain.
authors: M
authors: M
authors: Casanova
authors: M L
authors: ML
authors: Gil
authors: L
authors: L
authors: Cardeñoso
authors: J P
authors: JP
authors: Martinez
authors: R
authors: R
authors: Sentandreu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000946:Antigens, Fungal / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D002176:Candida albicans / Q000254:growth & development* / Q000276:immunology; D002473:Cell Wall / Q000276:immunology*; D000939:Epitopes / Q000276:immunology / Q000302:isolation & purification*; D006825:Hybridomas / Q000378:metabolism; D007106:Immune Sera; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D012100:Reproduction, Asexual
chemical_list: D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D000954:Antigens, Surface; D000939:Epitopes; D007106:Immune Sera; D008562:Membrane Glycoproteins; C025363:mannoproteins