Gastrodin interaction with human fibrinogen: Anticoagulant effects and binding studies

In an effort to identify the anticoagulant activity of gastrodin (GAS) and to investigate the possibility of its use as a novel anticoagulant drug, the binding characteristics of GAS to human fibrinogen (Fg) were studied by using a quartz crystal microbalance (QCM) biosensor, anticoagulant animal experiments, and a molecular docking simulation. Real-time kinetic analysis with the QCM biosensor revealed that the in vitro binding of GAS to Fg was strong under physiological ionic conditions as the determined equilibrium dissociation constant (K-D) was 1.94 x 10(-6) M. To check whether this strong binding may influence the natural coagulation function of Fg, the in vivo effect of GAS on the coagulation system of rats was examined. The results showed that GAS can significantly prolong the coagulation time (CT) and decrease the Fg content, while it had no effect on the activated kaolin partial thromboplastin time (KPTT) or prothrombin time (PT) in rats. To clarify the mechanism of the specific interaction, a molecular docking simulation was also performed to provide reasonable binding models for the interaction of GAS with Fg at the atomic level. GAS binds strongly to the inherent polymerization sites "a" and "b" (holes) on the Fg molecule with similar binding free energies of about -34 kJ mol(-1). Altogether, these findings confirmed first that GAS possesses anticoagulant activity and that the possible anticoagulation mechanism of GAS mainly involves its interference with the knob-to-hole interactions between fibrin molecules, thereby effectively inhibiting the formation of clots and decreasing the risk of thrombosis. The study has also shown the potential usefulness of QCM biosensor technology for the rapid screening of drug-protein interactions.Chemistry, MultidisciplinarySCI(E)EIPubMed9ARTICLE307807-78151