title: Antithrombotic activity of kininogen is mediated by inhibitory effects of domain 3 during arterial injury in vivo.
abstract: High-molecular-weight kininogen (HK) and its domain 3 (D3) exhibit anticoagulant properties and inhibit platelet activation at low thrombin concentration in vitro. We hypothesized that the rapid occlusive thrombosis in HK-deficient (HKd) rats following endothelial injury of the aorta results from enhanced platelet aggregation by thrombin. The effects of D3 (G235-M357) or D3-derived peptides on thrombosis in vivo were tested. D3 and its exon 7C terminal peptide (E7CP, K270-Q292), expressed as glutathione S-transferase (GST) fusion proteins (GST-D3, GST-E7CP), or GST alone, as well as cleaved HK (HKa) or synthetic peptide E7CP, were infused intravenously 10 min before endothelial injury. Blood flow was reduced down to 10% of baseline flow within 28 +/- 5.2 min by a platelet-fibrin thrombus in GST-treated HKd rats compared with >240 min in GST-treated normal HK rats (wild type). GST-D3, GST-E7CP, HKa, or E7CP infusion prolonged the flow time to 233, >240, 223, and >240 min, respectively, in HKd rats. When GST-E7CP was infused 10 min after the injury, blood flow was maintained for >240 min. Thrombin-antithrombin concentrations were elevated by injury in HKd rats receiving GST from 35 to 55 microg/l and decreased with GST-E7CP, HKa, or E7CP reconstitution to 40, 15, and 9 microg/l, respectively. We conclude that HKd rats are prothrombotic and that HKa, kininogen D3, and its fragment E7CP modulate arterial thrombosis after endothelial injury.
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple Univeristy School of Medicine, 3400 N. Broad St., Rm. 418 OMS, Philadelphia, PA 19140, USA.
authors.forename: Sarmina
authors.initials: S
authors.lastname: Hassan
authors.forename: Irma M
authors.initials: IM
authors.lastname: Sainz
authors.forename: Mohammad M
authors.initials: MM
authors.lastname: Khan
authors.forename: Harlan N
authors.initials: HN
authors.lastname: Bradford
authors.forename: Irma
authors.initials: I
authors.lastname: Isordia-Salas
authors.forename: Sakeen W
authors.initials: SW
authors.lastname: Kashem
authors.forename: R Balfour
authors.initials: RB
authors.lastname: Sartor
authors.forename: Robert W
authors.initials: RW
authors.lastname: Colman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D000990:Antithrombin III; D001011:Aorta / Q000187:drug effects / Q000293:injuries / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D001783:Blood Flow Velocity; D004195:Disease Models, Animal; D004730:Endothelium, Vascular / Q000187:drug effects / Q000293:injuries / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D005337:Fibrin / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use; D005982:Glutathione Transferase / Q000235:genetics; D019679:Kininogen, High-Molecular-Weight / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use; D008297:Male; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000097:blood; D010974:Platelet Aggregation; D017434:Protein Structure, Tertiary; D051381:Rats; D011917:Rats, Inbred Lew / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012039:Regional Blood Flow; D013917:Thrombin / Q000378:metabolism; D013927:Thrombosis / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control
chemical_list: D005343:Fibrinolytic Agents; D019679:Kininogen, High-Molecular-Weight; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C046193:antithrombin III-protease complex; D000990:Antithrombin III; D005337:Fibrin; D005982:Glutathione Transferase; D010447:Peptide Hydrolases; D013917:Thrombin