Two-dimensional electrophoresis, MALDI-TOF MS and blotting for identification and characterisation of pathogenesis-related proteins in wheat cultivar

摘要

Protein sets of different wheat cultivars were studied by two-dimensional polyacrylamide gel electrophoresis (2-D-PAGE). One of the wheat cultivars showed several differentially expressed spots, which were submitted to further investigation and identification. The protein spots interest (several spots of the differentially expressed proteins) were excised from the gel, and digested with trypsin. The digests after purification were analysed by MALDI-TOF MS and the proteins were identified by similarity searching in the protein database. Using this proteomic approach, one chitinase, one inhibitor and one heat shock protein were identified in the water- and salt-soluble extract of a wheat cultivar. PR-protein- related epitopes were found to be allergenic/cross-reactive determinant. This study demonstrated, that the sera of patients suffering from cereal allergy contained IgE anti- bodies which bound to some polypeptides of the globulin/albumin fraction of wheat flour. Our work describe an array of techniques suitable for characterisation and identification of components of a complex extract of wheat proteins from point of view of food safety and quality and and healthy nutrition.