β-Conglycinin is one of the major allergens existed in soybean.N-Glycans attached to theβ-conglycinin influenced the immunoreactivity and antigen presenting efficiency ofβ-conglycinin.In this study,we described a n...β-Conglycinin is one of the major allergens existed in soybean.N-Glycans attached to theβ-conglycinin influenced the immunoreactivity and antigen presenting efficiency ofβ-conglycinin.In this study,we described a new method used to release and collect the N-glycans fromβ-conglycinin,and the N-glycans existed in linear epitopes ofβ-conglycinin were identified.Glycopeptides hydrolyzed fromβ-conglycinin were purified by cotton hydrophilic chromatography.Trifluoromethylsulfonic acid was then used to release glycans from glycopeptides,and new glycopeptides containing one single N-acety1-D-glucosamine(G1 cNAc)moiety were then utilized for mass spectrometry.Five glycosylation sites(Asn-199,Asn-455,Asn-215,Asn-489 and Asn-326)and 22 kinds of glycopeptides were identified.It is noteworthy that the peptide VVN^(#)ATSNL(where^(#)represents for the glycosylation site)was analyzed to be both glycopeptide and linear epitope.Our results provided a new method for the N-glycoform analysis of food allergens,and laid a foundation for understanding the relationship between glyco sylation and food allergy.展开更多
基金funded by National Natural Science Foundation of China(31870798,31972024)Shaanxi Province Innovation Capability Support Plan-Science and Technology Innovation Team(2020TD-044)+2 种基金Key Laboratory of Glycobiology and Glycoengineering of Xi’an(2019219514SYS010CG032)Natural Science Project of Shaanxi Provincial Department of Education(21JK0929)Science and Technology Resources Sharing Platform Project of Science and Technology Department in Shaanxi Province(2022PT-46)。
文摘β-Conglycinin is one of the major allergens existed in soybean.N-Glycans attached to theβ-conglycinin influenced the immunoreactivity and antigen presenting efficiency ofβ-conglycinin.In this study,we described a new method used to release and collect the N-glycans fromβ-conglycinin,and the N-glycans existed in linear epitopes ofβ-conglycinin were identified.Glycopeptides hydrolyzed fromβ-conglycinin were purified by cotton hydrophilic chromatography.Trifluoromethylsulfonic acid was then used to release glycans from glycopeptides,and new glycopeptides containing one single N-acety1-D-glucosamine(G1 cNAc)moiety were then utilized for mass spectrometry.Five glycosylation sites(Asn-199,Asn-455,Asn-215,Asn-489 and Asn-326)and 22 kinds of glycopeptides were identified.It is noteworthy that the peptide VVN^(#)ATSNL(where^(#)represents for the glycosylation site)was analyzed to be both glycopeptide and linear epitope.Our results provided a new method for the N-glycoform analysis of food allergens,and laid a foundation for understanding the relationship between glyco sylation and food allergy.
