Magnesium Activates Microsecond Dynamics to Regulate Integrin-Collagen Recognition

Integrin receptors bind collagen via metal-mediated interactions that are modulated by magnesium (Mg²⁺) levels in the extracellular matrix. Nuclear magnetic resonance-based relaxation experiments, isothermal titration calorimetry, and adhesion assays reveal that Mg²⁺ functions as both a structural anchor and dynamic switch of the α₁β₁ integrin I domain (α₁I). Specifically, Mg²⁺ binding activates micro- to millisecond timescale motions of residues distal to the binding site, particularly those surrounding the salt bridge at helix 7 and near the metal ion-dependent adhesion site. Mutagenesis of these residues impacts α₁I functional activity, thereby suggesting that Mg-bound α₁I dynamics are important for collagen binding and consequent allosteric rearrangement of the low-affinity closed to high-affinity open conformation. We propose a multistep recognition mechanism for α₁I-Mg-collagen interactions involving both conformational selection and induced-fit processes. Our findings unravel the multifaceted role of Mg²⁺ in integrin-collagen recognition and assist in elucidating the molecular mechanisms by which metals regulate protein-protein interactions.