Energy landscape differences among integrins establish the framework for understanding activation

Why do integrins differ in basal activity, and how does affinity for soluble ligand correlate with cellular adhesiveness? We show that basal conformational equilibrium set points for integrin α₄β₁ are cell type specific and differ from integrin α₅β₁ when the two integrins are coexpressed on the same cell. Although α₄β₁ is easier to activate, its high-affinity state binds vascular cell adhesion molecule and fibronectin 100- to 1,000-fold more weakly than α₅β₁ binds fibronectin. Furthermore, the difference in affinity between the high- and low-affinity states is more compressed in α₄β₁ (600- to 800-fold) than in α₅β₁ (4,000- to 6,000-fold). α₄β₁ basal conformational equilibria differ among three cell types, define affinity for soluble ligand and readiness for priming, and may reflect differences in interactions with intracellular adaptors but do not predict cellular adhesiveness for immobilized ligand. The measurements here provide a necessary framework for understanding integrin activation in intact cells, including activation of integrin adhesiveness by application of tensile force by the cytoskeleton, across ligand-integrin-adaptor complexes.