Chapter title |
AFM-Based Single-Molecule Force Spectroscopy of Proteins
|
---|---|
Chapter number | 3 |
Book title |
Nanoscale Imaging
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8591-3_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8590-6, 978-1-4939-8591-3
|
Authors |
Zackary N. Scholl, Piotr E. Marszalek |
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. |
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