Mechanical unfolding of ensemble biomolecular structures by shear force |
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Authors: | Changpeng Hu Sagun Jonchhe Pravin Pokhrel Deepak Karna Hanbin Mao |
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Institution: | Department of Chemistry & Biochemistry and School of Biomedical Sciences, Advanced Materials and Liquid Crystal Institute, Kent State University Kent, OH 44242 USA, |
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Abstract: | Mechanical unfolding of biomolecular structures has been exclusively performed at the single-molecule level by single-molecule force spectroscopy (SMFS) techniques. Here we transformed sophisticated mechanical investigations on individual molecules into a simple platform suitable for molecular ensembles. By using shear flow inside a homogenizer tip, DNA secondary structures such as i-motifs are unfolded by shear force up to 50 pN at a 77 796 s−1 shear rate. We found that the larger the molecules, the higher the exerted shear forces. This shear force approach revealed affinity between ligands and i-motif structures. It also demonstrated a mechano-click reaction in which a Cu(i) catalyzed azide–alkyne cycloaddition was modulated by shear force. We anticipate that this ensemble force spectroscopy method can investigate intra- and inter-molecular interactions with the throughput, accuracy, and robustness unparalleled to those of SMFS methods.Shear force in a homogenizer mechanically unfolds an ensemble set of biomolecular structures. |
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