Single-Molecule Assay for Proteolytic Susceptibility: Force-Induced Collagen Destabilization
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Updated: Feb 14, 2026

Multiplexed Single-molecule Force Proteolysis Measurements Using Magnetic Tweezers
Published on: July 25, 2012
Michael W H Kirkness1, Nancy R Forde2
1Department of Molecular Biology and Biochemistry, Burnaby, British Columbia, Canada.
Abstract:
Force plays a key role in regulating dynamics of biomolecular structure and interactions, yet techniques are lacking to manipulate and continuously read out this response with high throughput. We present an enzymatic assay for force-dependent accessibility of structure that makes use of a wireless mini-radio centrifuge force microscope to provide a real-time readout of kinetics. The microscope is designed for ease of use, fits in a standard centrifuge bucket, and offers high-throughput, video-rate readout of individual proteolytic cleavage events. Proteolysis measurements on thousands of tethered collagen molecules show a load-enhanced trypsin sensitivity, indicating destabilization of the triple helix.
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