title: House dust mite-derived amylase: allergenicity and physicochemical characterization.
abstract: Amylase activity was found in extracts of both Dermatophagoides pteronyssinus whole mite (0.16 U/mg) and spent growth medium (0.01 U/mg) but not in unused growth medium. It was also detected in all extracts of house dust obtained from mattresses (n = 20; geometric mean, 1.95 U/gm) and in 18 extracts of dust obtained from lounge room carpets (n = 20; geometric mean, 0.54 U/gm). Although the origins of amylase in dust are unclear, enzyme activity correlated with mite counts (n = 40; r = 0.35; p less than 0.05) and Der p I concentrations (r = 0.41; p less than 0.01). Mite amylase was purified from spent growth medium by affinity chromatography, gel filtration, and chromatofocusing. It was physicochemically similar to mammalian amylase with regard to molecular weight (60,000), charge heterogeneity (isoelectric point, 5 to 7) and the capacity to bind to an organomercurial affinity matrix. The optimum pH for enzymatic activity was revealed to be 6.4. IgE immunoblot studies demonstrated that the enzyme was allergenic and that its expression was dependent on the integrity of intrachain disulfide bonds. Sera from 25% of mite-allergic children and 46% of mite-allergic adults contained specific IgE to mite amylase. IgE to amylase was associated (p less than 0.01) with increased concentrations of total mite-specific IgE determined with a direct RAST assay.
authors.affiliation: Western Australian Research Institute for Child Health, Princess Margaret Hospital, Perth.
authors.forename: F R
authors.initials: FR
authors.lastname: Lake
authors.forename: L D
authors.initials: LD
authors.lastname: Ward
authors.forename: R J
authors.initials: RJ
authors.lastname: Simpson
authors.forename: P J
authors.initials: PJ
authors.lastname: Thompson
authors.forename: G A
authors.initials: GA
authors.lastname: Stewart
mesh_terms: D000328:Adult; D000485:Allergens; D000681:Amylases / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D004391:Dust; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D008925:Mites / Q000201:enzymology*; D008970:Molecular Weight
chemical_list: D000485:Allergens; D004391:Dust; D007073:Immunoglobulin E; D000681:Amylases