Autonomous conformational regulation of β₃ integrin and the conformation-dependent property of HPA-1a alloantibodies.

Abstract

Integrin α/β heterodimer adopts a compact bent conformation in the resting state, and upon activation undergoes a large-scale conformational rearrangement. During the inside-out activation, signals impinging on the cytoplasmic tail of β subunit induce the α/β separation at the transmembrane and cytoplasmic domains, leading to the extended conformation of the ectodomain with the separated leg and the opening headpiece that is required for the high-affinity ligand binding. It remains enigmatic which integrin subunit drives the bent-to-extended conformational rearrangement in the inside-out activation. The β₃ integrins, including α_IIbβ₃ and α_Vβ₃, are the prototypes for understanding integrin structural regulation. The Leu33Pro polymorphism located at the β₃ PSI domain defines the human platelet-specific alloantigen (HPA) 1a/b, which provokes the alloimmune response leading to clinically important bleeding disorders. Some, but not all, anti-HPA-1a alloantibodies can distinguish the α_IIbβ₃ from α_Vβ₃ and affect their functions with unknown mechanisms. Here we designed a single-chain β₃ subunit that mimics a separation of α/β heterodimer on inside-out activation. Our crystallographic and functional studies show that the single-chain β₃ integrin folds into a bent conformation in solution but spontaneously extends on the cell surface. This demonstrates that the β₃ subunit autonomously drives the membrane-dependent conformational rearrangement during integrin activation. Using the single-chain β₃ integrin, we identified the conformation-dependent property of anti-HPA-1a alloantibodies, which enables them to differently recognize the β₃ in the bent state vs. the extended state and in the complex with α_IIb vs. α_V This study provides deeper understandings of integrin conformational activation on the cell surface.