Interaction of bovine factor XIIa with an inhibitor from bovine plasma.
An inhibitor of factor XIIa has been purified from bovine plasma and characterized (Thornton, R.D. and Kirby, E.P. (1987) J. Biol. Chem. 262, 12714-12721). This inhibitor interacts with XIIa to form a very stable complex with a 1:1 stoichiometry. The active site of XIIa, located on the light chain, is directly involved in the interaction, and complex formation between factor XIIa inhibitor and XIIa can be blocked by diisopropyl fluorophosphate, corn trypsin inhibitor, or the chromogenic substrate S2302. Incubation of the complex with excess XIIa does not result in cleavage of the complex. The complex does not spontaneously dissociate and is stable to boiling, SDS, thiocyanate, acid, and hydroxylamine or Tris at pH 7-10. In addition to complex formation, a cleaved form of factor XIIa inhibitor can be observed. We suggest that the inhibitor is acting as a mechanism-based inactivator, using the criteria of time-dependent inactivation under pseudo-first-order conditions, 1:1 stoichiometry, active site involvement, kinetic protection by substrate or by an active site inhibitor, and partitioning between cleavage of factor XIIa inhibitor and inactivation by complex formation.
Thornton
R D
RD
Department of Biochemistry, Temple University School of Medicine, Philadelphia, PA 19140.
Kirby
E P
EP
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002863:Chromogenic Compounds; D005174:Factor XII / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D015956:Factor XIIa; D007531:Isoflurophate / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D009842:Oligopeptides / Q000494:pharmacology; D011480:Protease Inhibitors / Q000097:blood* / Q000302:isolation & purification; D012697:Serine Endopeptidases / Q000302:isolation & purification; D015842:Serine Proteinase Inhibitors; D014361:Trypsin Inhibitors / Q000494:pharmacology
D002863:Chromogenic Compounds; D046911:Macromolecular Substances; D009842:Oligopeptides; D011480:Protease Inhibitors; D015842:Serine Proteinase Inhibitors; D014361:Trypsin Inhibitors; D007531:Isoflurophate; C026148:prolyl-phenylalanyl-arginine-4-nitroanilide; D005174:Factor XII; D012697:Serine Endopeptidases; D015956:Factor XIIa