Evidence for multiple enzyme site involvement in the modulation of thrombin activity by products of prothrombin proteolysis.
Kinetic evidence is presented for the interaction of prothrombin with several distinctive topological regions of the thrombin molecule. Modulations of thrombin catalytic activity on the protein substrates prothrombin and prethrombin 1 are demonstrated that involve the fragment 1 and fragment 2 portions. The inhibitory effects are demonstrably non-competitive. In addition to exhibiting non-competitive inhibition, fragment 2 is capable of enhancing proteolysis by thrombin; and therefore to react with a second region of the enzyme. On the basis of the crystallographic studies of the complex between fragment 2 and thrombin (Arni et al., Biochemistry 32 (1992) 4727), this activating site is proposed to be associated with exosite II. The allosteric switch between procoagulant and anticoagulant activities identified from studies by Di Cera (Dang et al., Proc. Natl. Acad. Sci USA 92 (1995) 5977) could be 'thrown' by a macromolecular effector that is generated during thrombin formation--a plausible mechanism for switching that deserves further investigation.
Shi
F
F
American Red Cross Blood Services, Detroit, MI 48232, USA.
Hogg
P J
PJ
Winzor
D J
DJ
Jackson
C M
CM
D000495:Allosteric Site; D020134:Catalytic Domain; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002863:Chromogenic Compounds / Q000378:metabolism; D004792:Enzyme Precursors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D009842:Oligopeptides / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011516:Prothrombin / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity; D013917:Thrombin / Q000737:chemistry* / Q000378:metabolism*
D002863:Chromogenic Compounds; D004792:Enzyme Precursors; D046911:Macromolecular Substances; D009842:Oligopeptides; D010446:Peptide Fragments; C035205:methylsulfonylleucyl-glycyl-arginine-4-nitroanilide; D011516:Prothrombin; C012007:prethrombins; D013917:Thrombin