Force-induced changes in protein structure and function underlie how cells respond to mechanical stress. However, existing methods for probing protein conformations under force are largely incompatible with biochemical and structural analyses such as electron microscopy. To overcome this limitation, we developed a DNA-based nanodevice that harnesses the well-defined geometry of DNA origami and the programmable mechanics of DNA hairpins to apply controlled forces to proteins.As a proof of concept, we investigated the R1–R2 segment of the talin1 rod domain, which consists of two α-helical bundles that reversibly unfold under tension to expose cryptic vinculin-binding sites. Electron microscopy confirmed tension-dependent extension of the protein, while biochemical assays demonstrated enhanced vinculin binding under load. Furthermore, the nanodevice was adapted for pull-down experiments with cell lysates, enabling the identification of force-dependent interactors of the target protein. Using this approach, we discovered filamins as previously unrecognized tension-dependent talin binders. Together, these results establish the DNA nanodevice as a versatile molecular toolbox for studying mechanosensitive proteins and mapping their force-dependent interaction networks.
Kim et al. (Sun,) studied this question.
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