Engineering Protein-Based Lipid-Binding Nanovesicles via Catechol-Amine-Derived Coacervation with Their Underlying...

The development of nonphospholipid nanovesicles has garnered tremendous attention as a viable alternative to traditional liposomal nanovesicles. Protein/peptide-based nanovesicles have demonstrated their potential to reduce immunogenicity while enhancing bioactivity. However, a fundamental understanding of how proteinaceous vesicles interact with lipids and cell membranes remains elusive. In this study, we engineered a series of protamine-based nonphospholipid nanovesicles by modulating intramolecular catechol-amine interactions. By grafting trihydroxybenzene (GA) and catechol (CA) groups onto the protamine (Prot), a salt-triggered coacervation was observed in an alkaline environment with the size of as-prepared vesicles ranging from 200 to 1200 nm. The bonding affinity to lipid interfaces followed the order of Prot-CA-Fe³⁺(25 μM) > Prot-CA-Fe³⁺(10 μM) > Prot-CA > original Prot with the underlying nanomechanics investigated by the lipid bubble force measurement. Direct quantification of interactions between the nanovesicles and living human gingival fibroblasts was performed by using surface charge difference mapping. Introducing trace amounts of Fe³⁺ (at 10 and 25 μM) enhanced vesicle-lipid interactions via the synergy of catechol-amine interactions and Fe³⁺-induced complexation. This work provides improved valuable insights into the interactions between nanovesicles and cell membranes, offering an energetic paradigm for modulating cell-target delivery processes via intramolecular short-range interactions.