Enzyme-Instructed Peptide Assembly Favored by Preorganization for Cancer Cell Membrane Engineering

Innovative methods for engineering cancer cell membranes promise to manipulate cell-cell interactions and boost cell-based cancer therapeutics. Here, we illustrate an in situ approach to selectively modify cancer cell membranes by employing an enzyme-instructed peptide self-assembly (EISA) strategy. Using three phosphopeptides (pY1, pY2, and pY3) targeting the membrane-bound epidermal growth factor receptor (EGFR) and differing in just one phosphorylated tyrosine, we reveal that site specific phosphorylation patterns in pY1, pY2, and pY3 can distinctly command their preorganization levels, self-assembling kinetics, and spatial distributions of the resultant peptide assemblies in cellulo. Overall, pY1 is the most capable of producing preorganized assemblies and shows the fastest dephosphorylation reaction in the presence of alkaline phosphatase (ALP), as well as the highest binding affinity for EGFR after dephosphorylation. Consequently, pY1 exhibits the greatest capacity to construct stable peptide assemblies on cancer cell membranes with the assistance of both ALP and EGFR. We further use peptide-protein and peptide- peptide co-assembly strategies to apply two types of antigens, namely ovalbumin (OVA) protein and dinitrophenyl (DNP) hapten respectively, on cancer cell membranes. This study demonstrates a very useful technique for the in situ construction of membrane bound peptide assemblies around cancer cells and implies a versatile strategy to artificially enrich cancer cell membrane components for potential cancer immunotherapy.

Automated summary

An enzyme-instructed peptide self-assembly (EISA) strategy is used to selectively modify cancer cell membranes, and different phosphorylation patterns in phosphopeptides targeting EGFR can impact their self-assembly abilities and binding affinity for EGFR. The phosphorylated peptide pY1 shows the highest capability for preorganization and the fastest dephosphorylation in the presence of alkaline phosphatase (ALP), making it the most effective at constructing stable peptide assemblies on cancer cell membranes. Additionally, peptide-protein and peptide-peptide co-assembly strategies are employed to apply antigens to cancer cell membranes, offering a potential technique for cancer immunotherapy.