Edited by Norma AllewellAntithrombin (AT) is an anticoagulant serpin that irreversibly inactivates the clotting proteinases factor Xa and thrombin by forming covalent complexes with them. Mutations in its critical domains, such as those that impair the conformational rearrangement required for proteinase inactivation, increase the risk of venous thrombosis. Á guila et al. characterize for the first time the destabilizing effects of mutations in the region of AT that makes contact with the proteinase in the final acyl-enzyme complex. Their work adds new insight into the unique structural intricacies of the inhibitory mechanism.Inhibitory serpins (serine proteinase inhibitors) belong to a superfamily of proteins with a characteristic fold of three  sheets, eight or nine ␣ helices, and a reactive center loop (RCL) 2 that contains a specific proteinase cleavage site (1). Of the ϳ1,500 identified serpin gene sequences, 36 are present in humans where they perform diverse functions. Antithrombin (AT), one of the 29 known human inhibitory serpins, is a major regulator of hemostasis, as inhibition of its targets thrombin and factor Xa (FXa) leads to a reduction in clotting. Inhibitory serpins are metastable: They undergo one of the largest known conformational changes upon forming covalent complexes with their proteinase targets (2). Whereas the importance of the RCL structure in this conformational change is well documented, much less is known about the structural requirements of residues in the base of the serpin, in particular the region that becomes the "landing place" for the inactivated proteinase in the covalent complex. Á guila et al. (3) now present clinical and biochemical proof that this region in the serpin is critical for inhibitory function and may be considered a new regulatory domain.Serpin action has been likened to that of a mousetrap. In the Michaelis complex, the proteinase is docked to the RCL of the native serpin; upon cleavage of the reactive site in the RCL, an acyl-enzyme intermediate is formed, and the N-terminal RCL fragment inserts as an extra strand into the principal  sheet of AT, rapidly translocating the attached proteinase 180°to the distal end of the serpin (Fig. 1, A and B). In this energetically favorable and stable complex, the proteinase active site is distorted, and the serpin acts as a suicide substrate (1, 2, 4, 5). Proteinase translocation, final docking, and distortion are steps in the inhibitory pathway of the branched serpin mechanism (Fig. 1C). Disturbing the serpin structure favors the alternative side of the branched mechanism, the substrate pathway, in which the acyl-enzyme intermediate completes hydrolysis to form a cleaved serpin and regenerated proteinase.The elaborate structure of inhibitory serpins makes them particularly prone to dysfunction caused by mutations. Moderate and even mild AT deficiency, as observed in patients with heterozygous mutations, significantly increases the risk of deep vein thrombosis and pulmonary embolism. In type II AT deficiency, normal a...