Papers Published
- Scholl, ZN; Marszalek, PE, AFM-Based Single-Molecule Force Spectroscopy of Proteins.,
Methods in Molecular Biology (Clifton, N.J.), vol. 1814
(January, 2018),
pp. 35-47 [doi] .
(last updated on 2023/06/01)Abstract:
Single-molecule force spectroscopy by AFM (AFM-SMFS) is an experimental methodology that allows unequivocal sensitivity and control for investigating and manipulating the mechanical properties of single molecules. The past 20 years of AFM-SMFS has provided numerous breakthroughs in the understanding of the mechanical properties and force-induced structural rearrangements of sugars, DNA, and proteins. Here, we focus on the application of AFM-SMFS to study proteins, since AFM-SMFS has succeeded in providing abundant information about protein folding pathways, kinetics, interactions, and misfolding. In this chapter we describe the experimental procedures for conducting a SMFS-AFM experiment-including purification of protein samples, setup and calibration of the AFM instrumentation, and the thorough and unbiased analysis of resulting AFM data.