Isolation and characterization of a major allergenic component (gp55) of Aspergillus fumigatus.
IgE class antibodies specific for antigens in a water-soluble extract of Aspergillus fumigatus (strain NHL-5759) were analyzed by immunoblotting with sera from patients with allergic bronchopulmonary aspergillosis. All the sera tested were reactive with a major 50 to 60 kd protein in the extract. This allergen, designated gp55, was purified by gel filtration and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigen was found to be present in the water-soluble extract in the form of a complex composed of approximately eight molecules of gp55. The carbohydrate and phosphate content of the purified antigen were 23.1% and 0.46%, respectively. The molar ratio of mannose to galactose residues was 2.76:1, and the protein was glycosylated predominantly with N-linked oligosaccharides. The serologic activity of the gp55 antigen was abolished by treatment with nonspecific protease (Pronase) but not by treatment with sodium metaperiodate or endoglycosidases. Thus the major antigenic site of the glycoprotein is located within its peptide moiety. The antigen itself displayed no chymotryptic or tryptic activity. The amino acid sequence of the 20 N-terminal residues of the antigen (ATPHEPVFFSWDAGAVTSFP) is different from that of any other protein previously reported.
Teshima
R
R
Division of Biochemistry and Immunochemistry, National Institute of Hygienic Sciences, Tokyo, Japan.
Ikebuchi
H
H
Sawada
J
J
Miyachi
S
S
Kitani
S
S
Iwama
M
M
Irie
M
M
Ichinoe
M
M
Terao
T
T
D000595:Amino Acid Sequence; D000908:Antibodies, Fungal / Q000097:blood; D000918:Antibody Specificity; D000946:Antigens, Fungal / Q000097:blood / Q000737:chemistry*; D001229:Aspergillosis, Allergic Bronchopulmonary / Q000097:blood; D001232:Aspergillus fumigatus / Q000276:immunology*; D002850:Chromatography, Gel; D010450:Endopeptidases / Q000378:metabolism; D005656:Fungal Proteins; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification*; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D005656:Fungal Proteins; D006023:Glycoproteins; C084249:gp55 protein, Aspergillus fumigatus; D007073:Immunoglobulin E; D010450:Endopeptidases