Candida albicans C3d receptor, isolated by using a monoclonal antibody.
Pseudohyphae of Candida albicans possess a receptor for C3d, a fragment of the complement component C3. This receptor was partially purified by using a monoclonal antibody (CA-A) that previously had been shown to inhibit the binding of C3d to C. albicans pseudohyphae. Purified immunoglobulin G from ascites fluid (CA-A) was coupled to a cyanogen bromide-activated Sepharose column, and an affinity-purified fraction (A2) from C. albicans pseudohyphae was obtained. This fraction inhibited rosetting of the EAC3d receptor by pseudohyphae and appeared to contain glycoprotein, since receptor activity could be removed when A2 was incubated with lectins specific for mannose and glucose. A2 was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and two polypeptides of approximately 60 and 70 kilodaltons (kDa) were consistently identified in reducing gels. The 60-kDa protein was identified as a glycoprotein by concanavalin A binding. A2 was further analyzed by high-pressure liquid chromatography (HPLC). Of three fractions obtained by HPLC, one containing the 60-kDa protein was found to have receptor activity. When analyzed by HPLC, this protein was found to contain mannose and glucose in approximately equal amounts. Both immunofluorescence and electron microscopy of pseudohyphae treated with CA-A identified A2 as a surface moiety. Thus, the C3d receptor of C. albicans, isolated with CA-A, is a glycoprotein of approximately 60 kDa.
Linehan
L
L
Department of Microbiology, University of Vermont, Burlington 05402.
Wadsworth
E
E
Calderone
R
R
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology; D002176:Candida albicans / Q000276:immunology*; D005656:Fungal Proteins / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D037102:Lectins; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D009504:Neutrophils / Q000276:immunology; D011951:Receptors, Complement / Q000276:immunology / Q000302:isolation & purification*; D017464:Receptors, Complement 3d
D000911:Antibodies, Monoclonal; D005656:Fungal Proteins; D037102:Lectins; D008562:Membrane Glycoproteins; D011951:Receptors, Complement; D017464:Receptors, Complement 3d