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Abstract

Biochemistry

High-Speed Magnetic Tweezers for Nanomechanical Measurements on Force-Sensitive Elements

Published: May 12th, 2023

DOI:

10.3791/65137

1Department of Physics, Pohang University of Science and Technology (POSTECH), 2School of Biological Sciences, Seoul National University, 3Institute for Molecular Biology and Genetics, Seoul National University, 4School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology (POSTECH)
* These authors contributed equally

Abstract

Single-molecule magnetic tweezers (MTs) have served as powerful tools to forcefully interrogate biomolecules, such as nucleic acids and proteins, and are therefore poised to be useful in the field of mechanobiology. Since the method commonly relies on image-based tracking of magnetic beads, the speed limit in recording and analyzing images, as well as the thermal fluctuations of the beads, has long hampered its application in observing small and fast structural changes in target molecules. This article describes detailed methods for the construction and operation of a high-resolution MT setup that can resolve nanoscale, millisecond dynamics of biomolecules and their complexes. As application examples, experiments with DNA hairpins and SNARE complexes (membrane-fusion machinery) are demonstrated, focusing on how their transient states and transitions can be detected in the presence of piconewton-scale forces. We expect that high-speed MTs will continue to enable high-precision nanomechanical measurements on molecules that sense, transmit, and generate forces in cells, and thereby deepen our molecular-level understanding of mechanobiology.

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Keywords Magnetic Tweezers

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