Platelet‐type von Willebrand disease: Local disorder of the platelet GPI GPI bα β‐switch drives high‐affinity binding to von Willebrand factor

摘要

Abstract Background Mutations in the β‐switch of GPI bα cause gain‐of‐function in the platelet‐type von Willebrand disease. Structures of free and A1‐bound GPI bα suggest that the β‐switch undergoes a conformational change from a coil to a β‐hairpin. Objectives Platelet‐type von Willebrand disease (VWD) mutations have been proposed to stabilize the β‐switch by shifting the equilibrium in favor of the β‐hairpin, a hypothesis predicated on the assumption that the complex crystal structure between A1 and GPI bα is the high‐affinity state. Methods Hydrogen‐deuterium exchange mass spectrometry is employed to test this hypothesis using G233V, M239V, G233V/M239V, W230L, and D235Y disease variants of GPI bα. If true, the expectation is a decrease in hydrogen‐deuterium exchange within the β‐switch as a result of newly formed hydrogen bonds between the β‐strands of the β‐hairpin. Results Hydrogen—exchange is enhanced, indicating that the β‐switch favors the disordered loop conformation. Hydrogen—exchange is corroborated by differential scanning calorimetry, which confirms that these mutations destabilize GPI bα by allowing the β‐switch to dissociate from the leucine‐rich‐repeat ( LRR ) domain. The stability of GPI bα and its A1 binding affinity, determined by surface plasmon resonance, are correlated to the extent of hydrogen exchange in the β‐switch. Conclusion These studies demonstrate that GPI bα with a disordered loop is binding‐competent and support a mechanism in which local disorder in the β‐switch exposes the LRR —domain of GPI bα enabling high‐affinity interactions with the A1 domain.