Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.7 (plasmin)
 9,023 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Glycoprotein IIb (GPIIb) and glycoprotein IIIa (GPIIIa) form a macromolecular complex on the activated platelet surface which contains the fibrinogen-binding site necessary for normal platelet aggregation. To identify the specific region of the fibrinogen molecule responsible for its interaction with the GPIIb-GPIIIa complex, purified fragment D1 (Mr = 100,000) and fragment E (Mr = 50,000) were prepared from plasmin digests of purified human fibrinogen. In addition, the polypeptide chain subunits A alpha, B beta, and gamma of fibrinogen were prepared. Using an enzyme-linked immunosorbent assay we have demonstrated that isolated fragment D1 in a solid phase system forms a complex with a mixture of GPIIb and GPIIIa. The binding of the GPIIb-GPIIIa mixture to fragment D1-coated plates reached saturation at 8 nM and to fibrinogen-coated plates at 24 nM. Isolated A alpha, B beta, and gamma chains were not reactive with added glycoproteins. Fragment E coated directly on plastic plates or immobilized on antibody-coated plastic plates did not form a complex with GPIIb-GPIIIa. Only fluid phase fibrinogen and fragment D1 but not fragment E were inhibitory toward formation of a complex between solid phase fibrinogen and GPIIb-GPIIIa. Isolated A alpha, B beta, and gamma chains at concentrations equivalent to fluid phase fibrinogen were inactive. Binding of fragment D1 but not fragment E to the GPIIb-GPIIIa complex was also demonstrated by rocket immunoelectrophoresis of the membrane glycoprotein mixture through a gel containing the individual fragments and subsequent autoradiography of the complex following exposure to 125I-anti-fibrinogen. These observations with isolated platelet membrane glycoproteins provide strong evidence that each of the D domains of the fibrinogen molecule interacts directly with the GPIIb-GPIIIa complex on the activated platelet surface, thus allowing formation of a tertiary molecular "bridge" across the surface of two adjacent activated platelets.
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PMID:Complex formation of platelet membrane glycoproteins IIb and IIIa with the fibrinogen D domain. 623 15

Platelets serve as a site for assembly of the proteins of the plasminogen activator system. Once bound to the platelet surface, tissue-type plasminogen activator manifests enhanced catalytic activity. Plasmin, once formed, also binds to the platelets surface and, at low concentrations, renders the platelet dysfunctional by cleaving glycoprotein IIIa selectively in the presence of bound fibrinogen. At higher concentrations (approximately 1 caseinolytic unit/ml), plasmin activates the platelet directly. Activated platelets also bind plasminogen and tissue-type plasminogen activator, and manifest enhanced catalytic efficiency of plasminogen activation. These observations suggest that plasminogen activation by tissue-type plasminogen activator is an autocatalytic process on the platelet surface, and that unique reciprocating mechanisms govern the interaction between platelets and the components of the plasminogen activator system.
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PMID:Platelets and plasminogen activation. 857 74

An activation-specific monoclonal antibody (MoAb) termed "Canine Activated Platelet 1" (CAP1) has been developed and partially characterized. Flow cytometric studies of isolated canine platelets, using adenosine diphosphate (ADP) and platelet activating factor (PAF) as agonists, demonstrated that CAPI binding site number was proportional to agonist strength and agonist concentration. MoAb CAP1 binding was diminished by ethylenediamine-tetraacetic acid, suggesting that the antigen was either stabilized by calcium or antigen binding to the platelet surface was mediated by calcium. ADP-activated gel-filtered platelets also demonstrated reduced binding of MoAB CAP1 even in the presence of 1 mM CaCl2. Binding of MoAb CAP1 could be partially restored by activating gel-filtered platelets with PAF, suggesting that the antigen was either present within platelet granule membranes or was exposed after binding of released proteins(s) with a platelet receptor. A monoclonal antibody to human platelet glycoprotein IIIa (GPIIIa), which cross-reacts with canine platelet GPIIIa regardless of platelet activation status, did not inhibit binding of MoAb CAP1. MoAb CAP1 bound to isolated canine fibrinogen captured on polystyrene microtiter plates in the absence of platelet proteins. Immunoblots indicated that MoAb CAP1 recognizes nonreduced fibrinogen as well as a plasmin digest of isolated canine fibrinogen. Results of the present studies suggest that MoAb CAP1 recognizes a receptor-induced binding site on canine fibrinogen.
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PMID:A platelet activation-specific monoclonal antibody that recognizes a receptor-induced binding site on canine fibrinogen. 881 40

Objective investigation of new inhibitors of blood protein or cellular systems that are activated during cardiopulmonary bypass (CPB) is impeded by the absence of a satisfactory animal model. Because most baboon hematologic proteins immunologically cross-react with those used for human assays, we developed a robust, reusable baboon model of CPB. Blood samples were obtained from adult baboons at six time intervals before, during, and after 60 minutes of partial CPB at 37 degrees C with peripheral cannulas. Both membrane (n = 7) and bubble oxygenators (n = 7) were investigated. We measured platelet and white blood cell counts; platelet response to adenosine diphosphate and release of beta-thromboglobulin; fibrinopeptide A, prothrombin fragment F1.2, thrombin-antithrombin complex, D-dimer, and plasmin-antiplasmin complex; activated complement (C3b/c and C4b/c); elastase-alpha1 proteinase inhibitor complex; and bleeding times. Adherent glycoprotein IIIa antigen in Triton X-100 washes of the perfusion circuit was also measured. Markers of baboon platelet, complement, and neutrophil activation and thrombosis significantly increased during CPB with bubble oxygenator systems but did not change appreciably in membrane oxygenator circuits. Markers of fibrinolysis, D-dimer, and plasmin-antiplasmin complex did not change with either oxygenator. The baboon model of CPB, when a bubble oxygenator is used, is a robust, reusable animal model for evaluating inhibitors of platelet, complement, and neutrophil activation and thrombosis during and after CPB.
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PMID:A baboon model for hematologic studies of cardiopulmonary bypass. 935 80