Role of arginine residues in the coagulant activity of high molecular weight kininogen.
High molecular weight (HMW) kininogen, the cofactor for activation of the contact system of plasma proteolysis, transports and optimally positions prekallikrein and factor XI on a negatively charged surface, allowing those zymogens to be activated by surface-bound factor XIIa. HMW kininogen circulates in plasma as a procofactor that, after cleavage by kallikrein or factor XIIa, gains ability to bind to the surface. The mechanism responsible for this increased affinity for the surface is unknown. We hypothesized that modification of arginine residues may prevent cleavage of HMW kininogen, since the initial kallikrein-induced cleavage sites on the HMW kininogen molecule are at the NH2 terminal and the COOH terminal of the bradykinin-containing portion of the molecule, each of which contains arginine. We found that modification with butanedione of four arginine residues in the HMW kininogen molecule prevented bradykinin release, which results from cleavage of HMW kininogen. Furthermore, HMW kininogen coagulant activity was lost, in proportion to the degree of arginine modification, until 6.6 residues had been modified. Complex formation with prekallikrein, however, was found to be uneffected by the modification of modified HMW kininogen. To account for the loss of coagulant activity, we also examined the ability of modified HMWKa (active cofactor) to bind to an activating surface. The affinity of modified HMWKa for kaolin was tenfold less than the affinity of unmodified HMWKa. These data suggest that arginine residues play a critical role in the ability of HMW kininogen to function as an activation cofactor, both by preventing the cleavages that produce HMWKa as well as by decreasing the affinity of HMWKa for the surface.
D001120:Arginine / Q000494:pharmacology*; D003029:Coagulants / Q000494:pharmacology*; D004852:Epoxy Compounds / Q000494:pharmacology; D006801:Humans; D007610:Kallikreins / Q000494:pharmacology; D007616:Kaolin / Q000378:metabolism; D007704:Kininogens / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D008970:Molecular Weight; D011288:Prekallikrein / Q000378:metabolism; D013329:Structure-Activity Relationship
D003029:Coagulants; D004852:Epoxy Compounds; D007704:Kininogens; D007616:Kaolin; C007366:diepoxybutane; D011288:Prekallikrein; D001120:Arginine; D007610:Kallikreins