ReviewThe Power of Force: Insights into the Protein Folding Process Using Single-Molecule Force Spectroscopy

•In this review, we have covered the latest topics in the field of protein folding using single-molecule force spectroscopy.•We include the main techniques used such as atomic force spectroscopy, optical tweezers, FRET, and magnetic tweezers.•We also cover the important advances in theoretical computational protein folding in complementing experimental results.•We have focused in advances made in the past five years or so, although we provide a good view of each technique beyond recent discoveries.•We include a section focused on the biological significance of using force to study protein folding and provide paradigmatic examples in biological systems where force seems to be a key element.•We also provide information regarding the most complex processes such as protein degradation of oxidative protein folding.•We finally provide some of the perspectives that the field of single-molecule force spectroscopy is heading toward.•We believe that the reader will be constructively walked through some of the most discussed topics in the field, providing also a good selection of references.•Overall, this review is directed toward a broad audience.

One of the major challenges in modern biophysics is observing and understanding conformational changes during complex molecular processes, from the fundamental protein folding to the function of molecular machines. Single-molecule techniques have been one of the major driving forces of the huge progress attained in the last few years. Recent advances in resolution of the experimental setups, aided by theoretical developments and molecular dynamics simulations, have revealed a much higher degree of complexity inside these molecular processes than previously reported using traditional ensemble measurements. This review sums up the evolution of these developments and gives an outlook on prospective discoveries.

Graphical AbstractDownload high-res image (85KB)Download full-size image