RETRACTED: Thrombin activation of factor XI on activated platelets requires the interaction of factor XI and platelet glycoprotein Ibα with thrombin anion-binding exosites I and II, respectively (Retracted Article. See vol 282, pg 29067, 2007)

Activation of factor XI (FXI) by thrombin on stimulated platelets plays a physiological role in hemostasis, providing additional thrombin generation required in cases of severe hemostatic challenge. Using a collection of 53 thrombin mutants, we identified 16 mutants with < 50% of the wild-type thrombin FXI-activating activity in the presence of dextran sulfate. These mutants mapped to anion-binding exosite (ABE) I, ABE-II, the Na+-binding site, and the 50-insertion loop. Only the ABE-II mutants showed reduced binding to dextran sulfate-linked agarose. Selected thrombin mutants in ABE-I (R68A, R70A, and R73A), ABE-II (R98A, R245A, and K248A), the 50-insertion loop (W50A), and the Na+-binding site ( E229A and R233A) with < 10% of the wildtype activity also showed a markedly reduced ability to activate FXI in the presence of stimulated platelets. The ABE-I, 50-insertion loop, and Na+-binding site mutants had impaired binding to FXI, but normal binding to glycocalicin, the soluble form of glycoprotein Ibalpha (GPIbalpha). In contrast, the ABE-II mutants were defective in binding to glycocalicin, but displayed normal binding to FXI. Our data support a quaternary complex model of thrombin activation of FXI on stimulated platelets. Thrombin bound to one GPIbalpha molecule, via ABE-II on its posterior surface, is properly oriented for its activation of FXI bound to a neighboring GPIalpha molecule, via ABE-I on its anterior surface. GPIbalpha plays a critical role in the co-localization of thrombin and FXI and the resultant efficient activation of FXI.