Protein Tethering for Folding Studies

Fatemeh Moayed; Roeland J van Wijk; David P Minde; Sander J Tans

doi:10.1007/978-1-4939-7271-5_3

Protein Tethering for Folding Studies

Methods Mol Biol. 2018:1665:43-51. doi: 10.1007/978-1-4939-7271-5_3.

Authors

Fatemeh Moayed¹, Roeland J van Wijk¹, David P Minde¹, Sander J Tans²

Affiliations

¹ AMOLF Institute, Science Park 104, 1098 XG, Amsterdam, The Netherlands.
² AMOLF Institute, Science Park 104, 1098 XG, Amsterdam, The Netherlands. tans@amolf.nl.

PMID: 28940063
DOI: 10.1007/978-1-4939-7271-5_3

Abstract

Optical tweezers allow the detection of unfolding and refolding transitions in individual proteins, and how interacting molecules such as chaperones affect these transitions. Typical methods that tether individual proteins are based on cysteine chemistry, which is less suitable for proteins with essential cysteines. Here we describe a cysteine-independent tethering protocol that can be performed in situ.

Keywords: Cysteine-independent linkages; In-situ tethering; Protein folding; Protein–DNA chimera; Single-molecule detection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Optical Tweezers*
Protein Conformation
Protein Denaturation
Protein Folding
Thermodynamics