Contribution of Chemical Modifications and Conformational Epitopes to IgE Binding by Ara h 3
AbstractRoasting is known to change the allergenic properties of peanuts. To study these observations at a molecular level, the relationship of IgE binding to the structure of Ara h 3 from raw and roasted peanuts was assessed. Ara h 3 (A3) was purified from raw (R), light roast (LR) and dark roast (DR) peanuts, the purity was assessed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the secondary structures were compared with circular dichroism (CD) spectroscopy. In order to understand the contribution of structure to IgE binding, the R A3 was partially denatured (PD) by heat treatment (65 °C for 2 h), subjected to CD spectroscopy and IgE spot blot analysis with sera from peanut- allergic individuals. While we observed that the secondary structure of purified A3 from R and LR peanut in solution was affected by the reduction of disulfide bonds and heat treatment when purified from the peanut following the roasting process, only small alterations were seen in the secondary structure. The purified LR A3 bound higher levels of IgE than the RA3. CD spectroscopy of PD A3 revealed a reduction in the percentage of alpha helices, and serum IgE binding. Therefore, while A3 purified from roasted peanuts did not show significant changes in secondary structure, it showed higher IgE binding than R A3. Therefore, the higher IgE binding to LR A3 was more likely to be due to chemical modifications than structural changes. However, a decrease in the IgE binding was seen if R A3 was deliberately unfolded, indicating that the structure played an important role in IgE binding to A3. View Full-Text
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Dyer, S.; Nesbit, J.B.; Cabanillas, B.; Cheng, H.; Hurlburt, B.K.; Maleki, S.J. Contribution of Chemical Modifications and Conformational Epitopes to IgE Binding by Ara h 3. Foods 2018, 7, 189.
Dyer S, Nesbit JB, Cabanillas B, Cheng H, Hurlburt BK, Maleki SJ. Contribution of Chemical Modifications and Conformational Epitopes to IgE Binding by Ara h 3. Foods. 2018; 7(11):189.Chicago/Turabian Style
Dyer, Scott; Nesbit, Jacqueline B.; Cabanillas, Beatriz; Cheng, Hsiaopo; Hurlburt, Barry K.; Maleki, Soheila J. 2018. "Contribution of Chemical Modifications and Conformational Epitopes to IgE Binding by Ara h 3." Foods 7, no. 11: 189.
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