Purification and identification of globulin-1 S allele as a novel allergen with N-glycans in wheat (Triticum aestivum)

Abstract

Wheat (Triticum aestivum) contains various allergenic proteins and induces diverse allergic reactions. Although many allergens are found to be glycoproteins, research on glycosylated allergens in wheat is rare. In this study, the main glycoprotein in wheat soluble protein was purified by sequential multi-column chromatography including affinity chromatography, gel filtration chromatography, and ion-exchange chromatography. The purified proteins were identified as globulin-1 S allele by liquid chromatography-tandem mass spectrometry and characterized by circular dichroism spectra. The strong IgE-binding capacity of the purified fractions was further demonstrated, suggesting globulin-1 S allele to be a novel allergen in wheat. Further de-N-glycosylated treatment by N-Glycosidase F showed that N-glycosylation enhanced the IgE-binding of globulin-1 S allele. These results identified globulin-1 S allele as a novel allergen in wheat, demonstrated the role of glycosylation in IgE-binding, and provided new insights into the prevention and treatment of wheat allergy.

Keywords: Allergens; Globulin-1 S allele; Glycoprotein; Wheat soluble protein.