title: The M358R variant of α(1)-proteinase inhibitor inhibits coagulation factor VIIa.
abstract: The naturally occurring M358R mutation of the plasma serpin α1-proteinase inhibitor (API) changes both its cleavable reactive centre bond to Arg-Ser and the efficacy with which it inhibits different proteases, reducing the rate of inhibition of neutrophil elastase, and enhancing that of thrombin, factor XIa, and kallikrein, by several orders of magnitude. Although another plasma serpin with an Arg-Ser reactive centre, antithrombin (AT), has been shown to inhibit factor VIIa (FVIIa), no published data are available with respect to FVIIa inhibition by API M358R. Recombinant bacterially-expressed API M358R and plasma-derived AT were therefore compared using gel-based and kinetic assays of FVIIa integrity and activity. Under pseudo-first order conditions of excess serpin over protease, both AT and API M358R formed denaturation-resistant inhibitory complexes with FVIIa in reactions accelerated by TF; AT, but not API M358R, also required heparin for maximal activity. The second order rate constant for heparin-independent API M358R-mediated FVIIa inhibition was determined to be 7.8 ± 0.8 × 10(2) M(-1)sec(-1). We conclude that API M358R inhibits FVIIa by forming inhibitory complexes of the serpin type more rapidly than AT in the absence of heparin. The likely 20-fold excess of API M358R over AT in patient plasma during inflammation raises the possibility that it could contribute to the hemorrhagic tendencies manifested by rare individuals expressing this mutant serpin.
authors.affiliation: Canadian Blood Services, Centre for Innovation, Hamilton, Ontario, Canada; Department of Pathology and Molecular Medicine†, McMaster University, Hamilton, Ontario, Canada. Electronic address: sheffiel@mcmaster.ca.
authors.forename: William P
authors.initials: WP
authors.lastname: Sheffield
authors.affiliation: Canadian Blood Services, Centre for Innovation, Hamilton, Ontario, Canada.
authors.forename: Varsha
authors.initials: V
authors.lastname: Bhakta
mesh_terms: D001777:Blood Coagulation / Q000502:physiology*; D015942:Factor VIIa / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D007700:Kinetics; D013329:Structure-Activity Relationship; D013925:Thromboplastin / Q000378:metabolism*; D000515:alpha 1-Antitrypsin / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000515:alpha 1-Antitrypsin; D013925:Thromboplastin; D015942:Factor VIIa
keywords: Alpha-1 proteinase inhibitor; Antithrombin; Blood coagulation; FVIIa; Heparin; Serpins