Optimization of Single-Molecule Assay for Proteolytic Susceptibility: Force-Induced Collagen Destabilization

Synopsis

Collagen has a triple helical structure, which allows for hierarchal assembly and tension resistance. Determining collagen’s structure under load is a complex problem; it has been a hotly debated topic with studies finding contradictory results. We use a centrifuge force microscope to perform single-molecule measurements and find that collagen’s triple helix is destabilized by an external load. We also report on our recent efforts towards a new surface chemistry based on an end-labelled block copolymer, F127 Pluronic - NHS. The surface chemistry is protein-free, reduces non-specific interactions with proteins, beads, quantum dots and peptides, is force-stable and is reproducible. The surface chemistry has a wide variety of uses in the fields of single-molecule force spectroscopy, lab-on-chip applications, and fluorescence microscopy.