Different Vinculin Binding Sites Use the Same Mechanism to Regulate Directional Force Transduction

• Published in: Biophysical journal Vol. 118; no. 6; pp. 1344 - 1356
• Main Authors: Kluger, Carleen, Braun, Lukas, Sedlak, Steffen M., Pippig, Diana A., Bauer, Magnus S., Miller, Ken, Milles, Lukas F., Gaub, Hermann E., Vogel, Viola
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 24-03-2020; The Biophysical Society
• Subjects: Binding Sites; Models, Molecular; Protein Binding; Talin - metabolism; Vinculin - metabolism
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/j.bpj.2019.12.042

Vinculin is a universal adaptor protein that transiently reinforces the mechanical stability of adhesion complexes. It stabilizes mechanical connections that cells establish between the actomyosin cytoskeleton and the extracellular matrix via integrins or to neighboring cells via cadherins, yet little is known regarding its mechanical design. Vinculin binding sites (VBSs) from different nonhomologous actin-binding proteins use conserved helical motifs to associate with the vinculin head domain. We studied the mechanical stability of such complexes by pulling VBS peptides derived from talin, α-actinin, and Shigella IpaA out of the vinculin head domain. Experimental data from atomic force microscopy single-molecule force spectroscopy and steered molecular dynamics (SMD) simulations both revealed greater mechanical stability of the complex for shear-like than for zipper-like pulling configurations. This suggests that reinforcement occurs along preferential force directions, thus stabilizing those cytoskeletal filament architectures that result in shear-like pulling geometries. Large force-induced conformational changes in the vinculin head domain, as well as protein-specific fine-tuning of the VBS sequence, including sequence inversion, allow for an even more nuanced force response.