Modulating ovalbumin-lipid bilayer interactions via fatty acid modification: A physicochemical characterization

Protein-lipid interactions are pivotal in various biological processes, including membrane trafficking, signal transduction, and cellular compartmentalization. This study investigates how fatty acid modification alters the physicochemical properties of ovalbumin (OVA) and its interactions with lipid bilayers, with implications for controlled release applications. We hypothesize that modifying OVA with different fatty acids will significantly alter its surface hydrophobicity and membrane affinity, thereby influencing its potential for targeted drug delivery. OVA was chemically modified with four fatty acids (myristic, palmitic, stearic, and oleic acids) using N-hydroxysuccinimide (NHS) esters. Surface hydrophobicity was assessed via 8-Anilino-1-naphthalenesulfonic acid ammonium (ANS) fluorescence assays, revealing that modifications increased hydrophobicity, particularly with longer fatty acid chains. Interactions with liposomes were characterized using dynamic light scattering (DLS) and zeta potential measurements, indicating enhanced liposome binding and altered bilayer properties due to fatty acid modifications. Quartz crystal microbalance with dissipation (QCM-D) analysis confirmed the insertion of fatty acid chains into the lipid bilayer. In vitro release experiments demonstrated that fatty acid modification significantly affected the release kinetics of OVA, with oleic acid-modified OVA showing the lowest cumulative release. Our findings highlight the role of fatty acid structure in modulating protein-lipid interactions and underscore the potential of fatty acid-modified OVA for developing targeted drug delivery systems. Future work will explore the underlying mechanisms and evaluate the therapeutic potential of these modified proteins in vivo.