Molecular Effects of Zwitterionic Peptide on Monolayer Lipid Membranes upon Enzyme-Catalyzed Degradation

Secretory phospholipase A₂ (sPLA₂), an enzyme overexpressed in numerous diseases, has been used to trigger structural transformations in lipid-based drug delivery systems, enabling payload release at target sites. Zwitterionic peptides are known for their superior antifouling properties, often outperforming poly(ethylene glycol) (PEG) surface modification by resisting protein adsorption. In this study, we examined lipid monolayers at the water–vapor interface on a Langmuir trough, incorporating varying molar fractions of zwitterionic peptide-conjugated lipids or PEG-conjugated lipids. Synchrotron X-ray surface techniques, including X-ray reflectivity and grazing incidence X-ray diffraction, were employed to analyze molecular packing, enzyme adsorption, enzyme-catalyzed lipid degradation, and metabolite reorganization at the interface, and microscopy was used to observe domain morphologies. The results demonstrate that zwitterionic peptides exhibit a significantly greater efficiency than PEG in stabilizing the interfacial monolayer packing structure against enzyme-catalyzed lipid degradation. However, contrary to most studies reporting strong resistance of zwitterionic materials to nonspecific protein absorption, enzyme absorption to the interface, which is interfacial and phospholipids specific, was not impeded by the presence of zwitterionic peptides at low molar ratios (≤10 mol %).