Unfolding and Refolding Proteins Using Single-Molecule AFM

Single-molecule atomic force microscopy (AFM) allows capturing the conformational dynamics of an individual molecule under force. In this chapter, we describe a protocol for conducting a protein nanomechanical experiment using AFM, covering both the force-extension and force-clamp modes. Combined, these experiments provide an integrated vista of the molecular mechanisms—and their associated kinetics—underpinning the mechanical unfolding and refolding of individual proteins when exposed to mechanical load.