title: Characterization of the 41kDa allergen Asp v 13, a subtilisin-like serine protease from Aspergillus versicolor.
abstract: Aspergillus versicolor is common on moldy building materials. Asp v 13, the principal allergen is produced by strains collected from across Canada. In this paper, we report a 1833bp Asp v 13 open reading frame predicted to encode a protein of 403 amino acids in length with three introns. A BLAST search of Asp v 13, a phylogenic tree calculation and alignment with its homologous proteins from other species indicated that Asp v 13 is a secretory, subtilisin-like serine protease widely distributed in Aspergillus species. His-tagged Asp v 13 was over-expressed in Escherichia coli and purified using Ni-NTA columns with a yield of 1mg/L. Based on immuno binding assay of recombinant protein both antibodies developed against the natural protein, and human sera IgE, the recombinant protein was similar to the natural form. Six IgE- and seven IgG-binding epitopes were also identified with selected human sera along the entire amino acid sequence of Asp v 13. Most residues binding these epitopes are exposed on the surface and correspond to charged regions of the molecule.
authors.affiliation: Department of Chemistry, Carleton University, Ottawa, ON, Canada.
authors.forename: C
authors.initials: C
authors.lastname: Shi
authors.forename: J D
authors.initials: JD
authors.lastname: Miller
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D000946:Antigens, Fungal / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001230:Aspergillus / Q000201:enzymology / Q000235:genetics* / Q000276:immunology*; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D057057:Serine Proteases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology
chemical_list: D000485:Allergens; D000946:Antigens, Fungal; D018985:Epitopes, B-Lymphocyte; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D057057:Serine Proteases