We are seeing patients in-person and through Video Visits. Learn more about how we’re keeping you safe and please review our updated visitor policy. Please also consider supporting Weill Cornell Medicine’s efforts to support our front-line workers.
Department of Anesthesiology

You are here

High-Speed Force Spectroscopy for Single Protein Unfolding.

TitleHigh-Speed Force Spectroscopy for Single Protein Unfolding.
Publication TypeJournal Article
Year of Publication2018
AuthorsSumbul F, Marchesi A, Takahashi H, Scheuring S, Rico F
JournalMethods Mol Biol
Volume1814
Pagination243-264
Date Published2018
ISSN1940-6029
Abstract

Single-molecule force spectroscopy (SMFS) measurements allow for quantification of the molecular forces required to unfold individual protein domains. Atomic force microscopy (AFM) is one of the long-established techniques for force spectroscopy (FS). Although FS at conventional AFM pulling rates provides valuable information on protein unfolding, in order to get a more complete picture of the mechanism, explore new regimes, and combine and compare experiments with simulations, we need higher pulling rates and μs-time resolution, now accessible via high-speed force spectroscopy (HS-FS). In this chapter, we provide a step-by-step protocol of HS-FS including sample preparation, measurements and analysis of the acquired data using HS-AFM with an illustrative example on unfolding of a well-studied concatamer made of eight repeats of the titin I91 domain.

DOI10.1007/978-1-4939-8591-3_15
Alternate JournalMethods Mol. Biol.
PubMed ID29956237