Molecular docking and molecular dynamics simulation of wheat gluten‐derived antioxidant peptides acting through the Keap1–Nrf2 pathway

Abstract BACKGROUND In our previous study, we successfully identified five peptides from wheat gluten: Ala‐Pro‐Ser‐Tyr (APSY), Leu‐Tyr (LY), Pro‐Tyr (PY), Arg‐Gly‐Gly‐Tyr (RGGY) and Tyr‐Gln (YQ). Molecular docking and molecular dynamics simulation methods were employed to investigate the interaction between these antioxidant peptides and the Kelch‐like ECH‐associated protein 1 (Keap1 protein), revealing the molecular mechanism of their non‐competitive binding. In addition, the total antioxidant capacity of the five peptides was determined using the 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt (ABTS) method. RESULTS The affinities of APSY, LY, PY, RGGY and YQ were −8.9, −8.3, −8.5, −9.1 and − 7.9 kcal mol −1 , respectively. The five peptides effectively bound to Keap1 protein through hydrogen, π‐σ, π‐alkyl and alkyl interactions. Significant roles were observed for the P1 pocket residue ARG‐415 and the P3 pocket residue ALA‐556 in the interactions of the Keap1–peptide complexes. Molecular dynamics simulations further elucidated the dynamic process of peptide binding to the Keap1 protein. All five peptides formed stable complexes with Keap1 protein, with van der Waals forces playing crucial roles in these complex systems, indicative of the peptides' strong binding ability to Keap1 protein. The van der Waals forces were −178.74, −123.11, −134.36, −132.59, and −121.44 kJ mol −1 for the Keap1–APSY, Keap1–LY, Keap1–PY, Keap1–RGGY and Keap1–YQ complexes, respectively. These peptides exhibited excellent antioxidant effects. Among them, the YQ peptide exhibited the highest total antioxidant capacity, with an activity value of 1.18 ± 0.06 mmol Trolox equivalent (TE) L −1 at a concentration of 0.10 mg mL −1 . The RGGY, PY, LY and APSY peptides followed in descending order, with activity values of 0.91 ± 0.05, 0.72 ± 0.06, 0.62 ± 0.04 and 0.60 ± 0.05 mmol TE L −1 , respectively. CONCLUSION These results unveiled the molecular mechanism by which the five antioxidant peptides act on active pockets through the Keap1–Nrf2 signaling pathway, providing a theoretical basis for the development of antioxidants. © 2024 Society of Chemical Industry.