PT - JOURNAL ARTICLE AU - Michael W.H. Kirkness AU - Nancy R. Forde TI - Single-molecule assay for proteolytic susceptibility using centrifuge force microscopy:force-induced destabilization of collagenā€˜s triple helix AID - 10.1101/171447 DP - 2017 Jan 01 TA - bioRxiv PG - 171447 4099 - http://biorxiv.org/content/early/2017/08/10/171447.short 4100 - http://biorxiv.org/content/early/2017/08/10/171447.full AB - 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 (MR.CFM) 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.