Comparison of proanthocyanidins A2 and B2 on IgE-reactivity and epitopes in Gly m 6 using multispectral, LC/MS-MS and molecular docking

This study comparatively analyzed the changes in IgE-reactivity and epitopes in proanthocyanidins A2- (PA-Gly m 6) and B2-Gly m 6 (PB-Gly m 6) conjugates prepared by alkali treatment at 80 degrees C for 20 min. Similar to the western blot, ELISA also showed a higher reduced IgE-reactivity in PA-Gly m 6 (70.12 %) than PB-Gly m 6 (63.17 %). SDS-PAGE demonstrated that proanthocyanidins A2 caused more formation of >180 kDa polymers than proanthocyanidins B2. Multispectral analyses revealed that PA-Gly m 6 exhibited more structural alteration (e.g., a decrease of & alpha;-helical content and ANS fluorescence intensity) to unfold protein structure than proanthocyanidins B2, improving the accessibility to modify Gly m 6 for shielding or destroying conformational epitopes. LC/MS-MS revealed that PA-Gly m 6 conjugates had a lower abundance of allergens, peptides and linear epitopes than PB-Gly m 6 conjugates. Molecular docking showed that proanthocyanidins A2 and B2 reacted with Gln-317 and Asn-94 of epitopes, respectively. Overall, proanthocyanidins A2 is more effective than proanthocyanidins B2 to decrease the IgE-reactivity of Gly m 6 due to more shielding or destruction of conformational epitopes and lower content allergens and linear epitopes, which was attributed to more proteincrosslinks formation and structural changes in PA-Gly m 6 conjugates.

Automated summary

This study comparatively analyzed the changes in IgE-reactivity and epitopes in proanthocyanidins A2- and B2-Gly m 6 conjugates prepared by alkali treatment. The results showed that PA-Gly m 6 had a higher reduced IgE-reactivity and caused more formation of >180 kDa polymers compared to PB-Gly m 6. PA-Gly m 6 exhibited more structural alteration and had a lower abundance of allergens, peptides, and linear epitopes than PB-Gly m 6. Overall, proanthocyanidins A2 were more effective in decreasing the IgE-reactivity of Gly m 6 due to more shielding or destruction of conformational epitopes and lower content of allergens and linear epitopes.