Immunological and chemical characterization of glycoproteins in IEF fractions of Blastomyces dermatitidis yeast lysate antigen.
After isoelectric focusing (IEF), fractions of a Blastomyces dermatitidis yeast lysate antigen were analysed for the presence of glycoproteins that may lead to cross-reactivity in immunoassays for the diagnosis of blastomycosis. Five major glycoproteins were apparent, two of which showed cross-reactivity when used in Western blots with sera obtained from dogs with histoplasmosis and coccidioidomycosis. These five glycoproteins were characterized for linkage to the proteins using N-glycosidase F (NGF) and for their lectin binding properties. The cross-reactive 235- and 160-kDa glycoproteins were found to possess mainly O-linked, high-mannose-type carbohydrates, and periodate-mediated oxidation of these molecules eliminated cross-reactivity observed with heterologous sera. Thus, the periodate-treated IEF antigens described here may be useful in solid-phase enzyme immunoassays for the diagnosis of blastomycosis.
Fisher
M A
MA
Department of Biological Sciences, Idaho State University, Pocatello 83209, USA.
Legendre
A M
AM
Scalarone
G M
GM
D000818:Animals; D000908:Antibodies, Fungal / Q000097:blood; D000946:Antigens, Fungal / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D001758:Blastomyces / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D001759:Blastomycosis / Q000097:blood / Q000276:immunology / Q000662:veterinary; D015153:Blotting, Western; D003047:Coccidioidomycosis / Q000097:blood / Q000276:immunology / Q000662:veterinary; D003429:Cross Reactions; D004283:Dog Diseases; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D006660:Histoplasmosis; D007525:Isoelectric Focusing; D008358:Mannose / Q000032:analysis; D008970:Molecular Weight
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D006023:Glycoproteins; D008358:Mannose