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

Abstract

Innovative methods for engineering cancer cell membranes promise to manipulate cell-cell interactions and boost cell-based cancer therapeutics. Here, we illustrate an <i>in situ</i> approach to selectively modify cancer cell membranes by employing an enzyme-instructed peptide self-assembly (EISA) strategy. Using three phosphopeptides (<b>pY1</b>, <b>pY2</b>, and <b>pY3</b>) targeting the membrane-bound epidermal growth factor receptor (EGFR) and differing in just one phosphorylated tyrosine, we reveal that site-specific phosphorylation patterns in <b>pY1</b>, <b>pY2</b>, and <b>pY3</b> can distinctly command their preorganization levels, self-assembling kinetics, and spatial distributions of the resultant peptide assemblies <i>in cellulo</i>. Overall, <b>pY1</b> 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, <b>pY1</b> 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 <i>in situ</i> 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.