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Determination of protein–protein interactions at the single-molecule level using optical tweezers

Published online by Cambridge University Press:  10 August 2022

Wendy N. Sánchez
Affiliation:
Biochemistry and Molecular Biology Department, Faculty of Chemistry and Pharmaceutical Sciences, Universidad de Chile, Santiago, Chile Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, Faculty of Science and Engineering, University of Groningen, Groningen, The Netherlands
Luka Robeson
Affiliation:
Biochemistry and Molecular Biology Department, Faculty of Chemistry and Pharmaceutical Sciences, Universidad de Chile, Santiago, Chile
Valentina Carrasco
Affiliation:
Biochemistry and Molecular Biology Department, Faculty of Chemistry and Pharmaceutical Sciences, Universidad de Chile, Santiago, Chile
Nataniel L. Figueroa
Affiliation:
Johannes Gutenberg-Universität Mainz, 55128 Mainz, Germany Helmholtz-Institut Mainz, GSI Helmholtzzentrum für Schwerionenforschung, 55128 Mainz, Germany
Francesca Burgos-Bravo
Affiliation:
Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720, USA
Christian A. M. Wilson*
Affiliation:
Biochemistry and Molecular Biology Department, Faculty of Chemistry and Pharmaceutical Sciences, Universidad de Chile, Santiago, Chile
Nathalie Casanova-Morales*
Affiliation:
Facultad de Artes Liberales, Universidad Adolfo Ibáñez, Santiago, Chile
*
Authors for correspondence: Christian A. M. Wilson, E-mail: yitowilson@gmail.com; Nathalie Casanova-Morales, E-mail: casa886@gmail.com
Authors for correspondence: Christian A. M. Wilson, E-mail: yitowilson@gmail.com; Nathalie Casanova-Morales, E-mail: casa886@gmail.com

Abstract

Biomolecular interactions are at the base of all physical processes within living organisms; the study of these interactions has led to the development of a plethora of different methods. Among these, single-molecule (in singulo) experiments have become relevant in recent years because these studies can give insight into mechanisms and interactions that are hidden for ensemble-based (in multiplo) methods. The focus of this review is on optical tweezer (OT) experiments, which can be used to apply and measure mechanical forces in molecular systems. OTs are based on optical trapping, where a laser is used to exert a force on a dielectric bead; and optically trap the bead at a controllable position in all three dimensions. Different experimental approaches have been developed to study protein–protein interactions using OTs, such as: (1) refolding and unfolding in trans interaction where one protein is tethered between the beads and the other protein is in the solution; (2) constant force in cis interaction where each protein is bound to a bead, and the tension is suddenly increased. The interaction may break after some time, giving information about the lifetime of the binding at that tension. And (3) force ramp in cis interaction where each protein is attached to a bead and a ramp force is applied until the interaction breaks. With these experiments, parameters such as kinetic constants (koff, kon), affinity values (KD), energy to the transition state ΔG, distance to the transition state Δx can be obtained. These parameters characterize the energy landscape of the interaction. Some parameters such as distance to the transition state can only be obtained from force spectroscopy experiments such as those described here.

Type
Review Article
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

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