Purification and partial characterization of the allergen Ag-54 from Cladosporium herbarum.
One important allergen, Ag-54, was extracted from Cladosporium herbarum and purified by a combination of diafiltration, gel filtrations and isoelectric focusing. The allergen, when examined by gel electrophoresis and high-performance liquid chromatography, was found to be essentially homogeneous with a molecular weight of about 20,000 daltons. Ag-54 is of glycoprotein nature. It contained 19.5% protein, calculated from its amino acid composition and 80.2% carbohydrate, quantified by methanolysis, trimethylsilyl derivatization and gas-liquid chromatography. The carbohydrate moiety was composed only of mannose, galactose and glucose in the ratio 1.0:0.6:1.3. Frequently occurring amino sugars like glucosamine and galactosamine could not be detected. Mass spectrometry of trimethylsilyl-derivatized methylglycosides demonstrated that sialic acid was not present.
D000485:Allergens; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000946:Antigens, Fungal / Q000302:isolation & purification*; D052179:Antigens, Plant; D002851:Chromatography, High Pressure Liquid; D002963:Cladosporium / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007122:Immunoelectrophoresis; D003904:Mitosporic Fungi / Q000276:immunology*; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid; D011817:Rabbits / Q000276:immunology; D012794:Sialic Acids / Q000032:analysis; D012967:Sodium Dodecyl Sulfate
D000485:Allergens; D000596:Amino Acids; D000946:Antigens, Fungal; D052179:Antigens, Plant; C046933:Cla h II protein, Cladosporium herbarum; D005656:Fungal Proteins; D007106:Immune Sera; D012794:Sialic Acids; D012967:Sodium Dodecyl Sulfate; D019158:N-Acetylneuraminic Acid