title: Murine thrombin lacks Na+ activation but retains high catalytic activity.
abstract: Human thrombin utilizes Na+ as a driving force for the cleavage of substrates mediating its procoagulant, prothrombotic, and signaling functions. Murine thrombin has Asp-222 in the Na+ binding site of the human enzyme replaced by Lys. The charge reversal substitution abrogates Na+ activation, which is partially restored with the K222D mutation, and ensures high activity even in the absence of Na+. This property makes the murine enzyme more resistant to the effect of mutations that destabilize Na+ binding and shift thrombin to its anticoagulant slow form. Compared with the human enzyme, murine thrombin cleaves fibrinogen and protein C with similar k(cat)/K(m) values but activates PAR1 and PAR4 with k(cat)/K(m) values 4- and 26-fold higher, respectively. The significantly higher specificity constant toward PAR4 accounts for the dominant role of this receptor in platelet activation in the mouse. Murine thrombin can also cleave substrates carrying Phe at P1, which potentially broadens the repertoire of molecular targets available to the enzyme in vivo.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Bush
authors.forename: Ryan W
authors.initials: RW
authors.lastname: Nelson
authors.forename: Enrico
authors.initials: E
authors.lastname: Di Cera
mesh_terms: D000818:Animals; D001665:Binding Sites; D002384:Catalysis; D003029:Coagulants / Q000737:chemistry*; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000737:chemistry; D005340:Fibrinogen / Q000737:chemistry; D005822:Genetic Vectors; D006801:Humans; D006868:Hydrolysis; D007668:Kidney / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D009154:Mutation; D015539:Platelet Activation; D011485:Protein Binding; D011486:Protein C / Q000737:chemistry; D012964:Sodium / Q000737:chemistry; D013917:Thrombin / Q000737:chemistry*
chemical_list: D003029:Coagulants; D011486:Protein C; D005340:Fibrinogen; D012964:Sodium; D013917:Thrombin