Spatiotemporal Resolved Live Cell Membrane Tracking through Photo‐click Reactions Enriched in Lipid Phase

A set of photo‐switchable monopeptides derived from cis‐β‐dibenzodiazocine‐l‐alanine (cis‐DBDAA) have been designed and synthesized, which are capable of photo‐click reacting with diaryltetrazoles or diarylsydnones in a hydrophobic phospholipid bilayer environment. The DBDAA monopeptides include both a hydrophobic tail on C‐terminal, providing high affinity toward lipid membrane, and a modularized functional moiety on N‐terminal, enabling rapid optimization of the self‐assembly strength to form multifunctional supramolecules. With the cis‐DBDAA monopeptides photo‐switched into trans‐configuration, we were able to disrupt the supramolecular assembly through an efficient photo‐click reaction across the lipid bilayer of liposomes. We reveal that the performance of the photo‐click reactions between the monopeptides and photo‐generated nitrile imine intermediates is significantly enhanced by enrichment of both reactants in the hydrophobic membrane lamel of liposomes. Enrichment of the DBDAA monopeptide in lipid phase serves as a convenient method to introduce bioorthogonal chemical handles on live cell membranes, which enables fluorescence labelling of single cell's membrane with high spatiotemporal resolution to facilitate the studies on cell membrane dynamics. A lipid phase mediated photo‐click cycloaddition between photo‐generated nitrile imines and energy‐storable dibenzo[c,g][1,2]diazocine (DBDA) was established, including optimized biocompatibility and highly spatiotemporal resolution as well. A supramolecular assembly of the photo‐switchable DBDA monopeptides was also exploited to implant the DBDA moiety on live cell membranes for studying their dynamics via the photo‐click reaction by using digital micromirror device (DMD) chip.