[2Ap] Nutiritions: Protein, Carbohydrate, Lipid, Vitamin, and Mineral

[2Ap-08] Glutathione and the Structure of Rice Allergen Proteins

*Yoshihiko Tokuji¹, Takako Tenmadate¹ (1. Obihiro University of Agriculture and Veterinary Medicine)

Keywords:rice, metabolome, protein, glutathione

Purpose: The study investigates the inhibitory effect of the rice variety Yukihikari on allergy. Despite reports suggesting intestinal fermentation, microflora, and permeability might contribute to this effect, the precise mechanism and causative agent remain unclear. The research aims to identify the characteristic components of Yukihikari through metabolome analysis and protein pattern comparison.Methods: Water-soluble and fat-soluble substances were extracted from Yukihikari rice, known for its allergy suppression, and from a non-suppressive variety. Metabolomic analysis was conducted using CE-MS for water-soluble substances and LC-MS for fat-soluble substances. Proteins were compared using Native PAGE, and differing protein bands were identified by LC-MS/MS. Antipeptide antibodies based on the identified proteins were used for Western analysis.Results: Metabolome Analysis: Yukihikari's glutathione content was found to be only 1/20th of the non-suppressive variety.Protein Comparison: Native PAGE showed a 350 kDa band in Yukihikari, which decreased with added glutathione.LC-MS/MS Analysis: The 350 kDa band included known allergenic proteins and members of the α-amylase/trypsin inhibitor family, rich in cysteine residues, suggesting glutathione's role in regulating disulfide bonds.Western Analysis: Antipeptide antibodies revealed the presence of a 350 kDa band, indicating that several constituent proteins form a complex.