IDENTIFICATION AND CHARACTERIZATION OF WHEAT-GRAIN ALBUMIN GLOBULIN ALLERGENS/

Bakers' asthma, an immediate-type allergic response to the inhalation of cereal flours, is an important occupational disease among workers o f the baking and milling industries, and the salt-soluble proteins of wheat and rye flour dust are considered the most relevant allergens. I n order to identify and characterize the major IgE-binding proteins, t he polypeptide composition of the albumin/globulin protein fraction ob tained from different cultivars was analyzed by sodium dodecyl sulfate -polyacrylamide gel electrophoresis (SDS-PAGE), and high-resolution tw o-dimensional polyacrylamide gel electrophoresis with immobilized pH g radients in the first dimension (IPG-Dalt), followed by immunoblotting with sera from asthmatic bakers. Relevant allergens were isolated by micropreparative IPG-Dalt and blotting onto polyvinylidenedifluoride m embranes and identified by amino acid composition analysis or N-termin al amino acid sequence analysis. SDS-PAGE, IPG-Dalt, and immunoblottin g demonstrated that the sera of the bakers allergic to flour contained IgE antibodies which bound to numerous albumin/globulin polypeptides in the 70, 55, 35, 26-28, and 14-15 kDa areas. More detailed investiga tions using IPG-Dalt revealed cultivar-specific differences in IgE-bin ding. It was also demonstrated that the majority of the allergens were not single polypeptide spots, but consisted of up to ten isoforms of similar molecular mass but different isoelectric points. Amino acid co mposition analysis and N-terminal amino acid sequence analysis, which were performed for nine allergens located in the 14-18, 26-28, and 35 kDa areas, revealed homologies to amylase/protease inhibitors, acyl-Co A oxidase and fructose-bisphosphate-aldolase from wheat, barley, maize , and rice, respectively.