title: Characterization of an antigenic chitosanase from the cellulolytic fungus Chaetomium globosum.
abstract: We are interested in identifying human fungal allergens and antigens from species common on water-damaged or damp building materials for use as marker proteins and diagnostic tests. The cellulolytic fungus Chaetomium globosum is common on damp materials in the building environment worldwide. ELISA and immunoblotting tests identified two related proteins of molecular weights 45 and 47 kDa which were identified as fungal antigens found on spore surfaces and in culture filtrate. The sequences were determined by liquid chromatography tandem mass spectrometry (LC-MS/MS), which indicated that the two proteins were chitosanases, confirmed by enzyme assay. The 47 kDa protein was not glycosylated and had an acidic pI of 4.5. These proteins have not been reported from other fungi and similar antigens were not seen in other fungi common in buildings. The production of polyclonal antibodies in rabbits showed the antigenicity of the target proteins and confirmed they were not artifacts of the isolation process. The proteins isolated are useful biomarkers for the detection of C. globosum in the building environment.
authors.affiliation: Ottawa-Carleton Institute of Chemistry, Carleton University, Ottawa, ON, Canada.
authors.forename: Natacha B
authors.initials: NB
authors.lastname: Provost
authors.forename: Chunhua
authors.initials: C
authors.lastname: Shi
authors.forename: Yi-Min
authors.initials: YM
authors.lastname: She
authors.forename: Terry D
authors.initials: TD
authors.lastname: Cyr
authors.forename: J David
authors.initials: JD
authors.lastname: Miller
mesh_terms: D000818:Animals; D000946:Antigens, Fungal / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*; D002597:Chaetomium / Q000201:enzymology* / Q000276:immunology* / Q000302:isolation & purification; D002853:Chromatography, Liquid; D004783:Environmental Microbiology; D004797:Enzyme-Linked Immunosorbent Assay; D006026:Glycoside Hydrolases / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007526:Isoelectric Point; D008970:Molecular Weight; D011817:Rabbits; D053719:Tandem Mass Spectrometry
chemical_list: D000946:Antigens, Fungal; D006026:Glycoside Hydrolases; C019868:chitosanase