Biochemistry and molecular biology of the main diagnostic antigen of Paracoccidioides brasiliensis.
The 43,000 dalton glycoprotein of Paracoccidioides brasiliensis (gp 43) is the main exocellular antigen recognized by sera from patients with paracoccidioidomycosis in a variety of serological assays. Specific conformational peptide epitopes are recognized by the human antibodies as determined by antigen deglycosylation. Procedures for the purification of the gp43 using immunoaffinity chromatography have been described. The secretion of the gp43 as a function of the growth curve, its partial aggregation with a proteolytic enzyme, ability to bind laminin, as well as to form circulating immunocomplexes in vivo could play a role in pathogenesis. Crude antigenic preparations depleted of gp43 epitopes lost their ability to elicit positive skin tests. Accordingly, the purified gp43 molecule induced delayed hypersensitivity reactions in man and infected animals, caused a T-CD4-dependent proliferation of lymph node cells from mice immunized with it, and of peripheral blood lymphocytes from an individual sensitized to P. brasiliensis by prolonged contact with the fungus. To identify the immunodominant epitopes in both humoral and cellular reactions, the gp43 gene has been cloned, sequenced, and partly expressed. It bears peptide sequences homologous to those of beta-1,3-glucanases from Candida albicans and Saccharomyces cerevisiae but has no enzymatic activity itself. The molecular weight of the unglycosylated antigen is 42,227. A single N-linked oligosaccharide chain in the gp43 contains alpha-D-mannopyranosyl, beta-D-galactofuranosyl and N-acetylglucosaminyl units with the predominant ratio of 10:2:2, and characteristics of a high mannose type.
Travassos
L R
LR
Universidade Federal de São Paulo, Escola Paulista de Medicina, Brasil.
Puccia
R
R
Cisalpino
P
P
Taborda
C
C
Rodrigues
E G
EG
Rodrigues
M
M
Silveira
J F
JF
Almeida
I C
IC
D000595:Amino Acid Sequence; D000818:Animals; D000946:Antigens, Fungal / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010228:Paracoccidioides / Q000276:immunology*; D010229:Paracoccidioidomycosis / Q000175:diagnosis*; D017386:Sequence Homology, Amino Acid
D000946:Antigens, Fungal; D006023:Glycoproteins