Protein engineering thrombin for optimal specificity and potency of anticoagulant activity in vivo.

摘要: Previous alanine scanning mutagenesis of thrombin revealed that substitution residues W50, K52, E229, and R233 (W60d, K60f, E217, R221 in chymotrypsinogen numbering) with altered the substrate specificity to favor anticoagulant protein C. Saturation mutagenesis, which were each substituted all 19 naturally occurring amino acids, resulted identification a single mutation, E229K, shifted by 130-fold activation C over procoagulant fibrinogen. E229K was also less effective activating platelets (18-fold), resistant inhibition antithrombin III (33-fold 22-fold presence absence heparin), displayed prolonged half-life plasma vitro (26-fold). Thus an optimal phenotype function as potent specific activator endogenous an...