Structural insights into integrin α₅β₁ opening by fibronectin ligand.

Abstract

Integrin α₅β₁ is a major fibronectin receptor critical for cell migration. Upon complex formation, fibronectin and α₅β₁ undergo conformational changes. While this is key for cell-tissue connections, its mechanism is unknown. Here, we report cryo-electron microscopy structures of native human α₅β₁ with fibronectin to 3.1-angstrom resolution, and in its resting state to 4.6-angstrom resolution. The α₅β₁-fibronectin complex revealed simultaneous interactions at the arginine-glycine-aspartate loop, the synergy site, and a newly identified binding site proximal to adjacent to metal ion-dependent adhesion site, inducing the translocation of helix α1 to secure integrin opening. Resting α₅β₁ adopts an incompletely bent conformation, challenging the model of integrin sharp bending inhibiting ligand binding. Our biochemical and structural analyses showed that affinity of α₅β₁ for fibronectin is increased with manganese ions (Mn²⁺) while adopting the half-bent conformation, indicating that ligand-binding affinity does not depend on conformation, and α₅β₁ opening is induced by ligand-binding.