Use of N-benzoyl-phenylalanyl-valyl-arginine p-nitroanilide and hirudin for the detection of enzymatic activity under prothrombin acylation by citraconic anhydride

Acylation of bovine prothrombin by cytraconic anhydride results in the formation of the thrombin active site, which provides the degradation of highly specific thrombin substrate, N-bensoyl-Phe-Val-Arg p-nitroanilide. This reaction is inhibited with hirudin, highly specific thrombin inhibitor.