Undigestible Gliadin Peptide Nanoparticles Penetrate Mucus and Reduce Mucus Production Driven by Intestinal Epithelial Cell Damage

Wheat protein is the most consumed plant protein in our diet, and there is an increased prevalence of wheat/gluten intolerance and adherence to a gluten-free diet in many countries. Despite the known immunodominant effect of undigested gliadin peptides responsible for gluten-related intolerance, it remains unclear if and how gliadin peptides self-assemble into ordered nanostructures during gastrointestinal digestion, as well as their biological impact on the mucus barrier function. In this study, we purified undigestible gliadin peptide nanoparticles (UGPNs) by ultracentrifugation and characterized their structural and physiochemical properties. The results demonstrate that the UGPNs are self-assembled nanostructures generated by cationic amino acids (Lys and Arg)-capped surfactant-like peptides (SLPs), mainly derived from γ-gliadin and α-gliadin. SLPs trigger the concentration-dependent self-assembly driven by β-sheet conformational transitions above their critical aggregation concentration (cac, ∼0.1 mg/mL). UGPNs can easily penetrate the mucus layer in Caco-2/HT29-MTX cocultures with a high P_app value (∼5.7 × 10^-6 cm/s) and reduce the production and thickness of the mucus layer driven by intestinal epithelial cell damage. Isothermal titration calorimetry and Langmuir monolayer studies indicate that the self-assembled state of UGPNs significantly affects their binding to DPPC/DOPE lipid membrane models. These results highlight the relevance of the self-assembly of gliadin peptides as a trigger of mucosal inflammation-related wheat/gluten intolerance.