Baxter, NJ, Zacharchenko, T, Barsukov, IL et al. (1 more author) (2017) Pressure-Dependent Chemical Shifts in the R3 Domain of Talin Show that It Is Thermodynamically Poised for Binding to Either Vinculin or RIAM. Structure, 25 (12). 1856-1866.e2. ISSN 0969-2126
Abstract
Talin mediates attachment of the cell to the extracellular matrix. It is targeted by the Rap1 effector RIAM to focal adhesion sites and subsequently undergoes force-induced conformational opening to recruit the actin-interacting protein vinculin. The conformational switch involves the talin R3 domain, which binds RIAM when closed and vinculin when open. Here, we apply pressure to R3 and measure ¹H, ¹⁵N, and ¹³C chemical shift changes, which are fitted using a simple model, and indicate that R3 is only 50% closed: the closed form is a four-helix bundle, while in the open state helix 1 is twisted out. Strikingly, a mutant of R3 that binds RIAM with an affinity similar to wild-type but more weakly to vinculin is shown to be 0.84 kJ mol‾¹ more stable when closed. These results demonstrate that R3 is thermodynamically poised to bind either RIAM or vinculin, and thus constitutes a good mechanosensitive switch.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Keywords: | hydrostatic pressure; cell adhesion; talin; focal adhesion complex; vinculin; singular value decomposition; chemical shift |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 24 Nov 2017 11:18 |
Last Modified: | 10 Apr 2018 08:57 |
Status: | Published |
Publisher: | Elsevier (Cell Press) |
Identification Number: | 10.1016/j.str.2017.10.008 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:124464 |