Focal adhesions are multiprotein complexes that couple actin to the extracellular matrix (ECM) and govern force transmission and signaling, thereby controlling cell migration, mechanotransduction, and invasion. Focal adhesions are primarily governed by the mechanosensitive vinculin-talin interface, yet actionable molecular levers to tune this system remain limited. We hypothesize that introducing hydrocarbon-stapled talin mimetics can selectively engage vinculin, bias its conformational ensemble, and stabilize adhesions in a migration-suppressing state. We have started quantifying vinculin’s engagement with surface plasmon resonance (SPR) to test this hypothesis and establish kinetic parameters for stapled talin-mimetics. We are currently performing competitive binding assays with native talin to evaluate displacement and constraints on native interaction, which will indicate conformational bias. We will perform a complementary X-ray crystallography of vinculin-peptide complexes to resolve peptide-enforced binding poses and conformational rearrangements to explain this mechanism holistically. Finally, we will assess functional implications in melanoma cells using adhesion assays to demonstrate that the peptide’s engagement stabilizes focal adhesions in a unique conformation that suppresses invasion. This work will offer a reproducible and generalizable mechanism for modulating force-transducing scaffolds to inform anti-metastatic approaches to cancer.
Essuman et al. (Sun,) studied this question.