Cross-Linking of Alpha 2-Plasmin Inhibitor to Fibrin by Fibrin- Stabilizing Factor

Total Page:16

File Type:pdf, Size:1020Kb

Cross-Linking of Alpha 2-Plasmin Inhibitor to Fibrin by Fibrin- Stabilizing Factor Cross-linking of alpha 2-plasmin inhibitor to fibrin by fibrin- stabilizing factor. Y Sakata, N Aoki J Clin Invest. 1980;65(2):290-297. https://doi.org/10.1172/JCI109671. Research Article The concentration of alpha 2-plasmin inhibitor in blood plasma is higher than that in serum obtained from the blood clotted in the presence of calcium ions, but is the same as that in serum obtained in the absence of calcium ions. Radiolabeled alpha2-plasmin inhibitor was covalently bound to fibrin only when calcium ions were present at the time of clotting of plasma or fibrinogen. Whereas, when batroxobin, a snake venom enzyme that lacks the ability to activate fibrin- stabilizing factor, was used for clotting fibrinogen, the binding was not observed. When fibrin-stablizing, factor-deficient plasma was clotted, the specific binding of alpha 2-plasmin inhibitor to fibrin did not occur even in the presence of calcium ions and the concentration of alpha 2-plasmin inhibitor in serum was the same as that in plasma. Monodansyl cadaverine, a fluorescent substrate of the fibrin-stablizing factor, was incorporated into alpha 2-plasmin inhibitor by activated fibrin-stablizing factor. All these findings indicate that alpha 2-plasmin inhibitor is cross-linked to fibrin by activated fibrin-stabilizing factor when blood is clotted. Analysis of alpha 2-plasmin inhibitor-incorporated fibrin by sodium dodecyl sulfate gel electrophoresis showed that the inhibitor was mainly cross-linked to polymerized alpha-chains of cross-linked fibrin. Cross-linking of alpha 2-plasmin inhibitor to fibrin renders fibrin clot less susceptible to fibrinolysis by plasmin. Find the latest version: https://jci.me/109671/pdf Cross-Linking of a2-Plasmin Inhibitor to Fibrin by Fibrin-stabilizing Factor YOICHI SAKATA and NOBuo AoKI, Institute of Hematology and Department of Medicine, Jichi Medical School, Tochigi-Ken, Japan 329-04 A B S TR A C T The concentration of a2-plasmin inhib- solution of fibrin (fibrinolysis) is believed to be checked itor in blood plasma is higher than that in serum ob- or retarded by natural inhibitors of fibrinolysis. tained from the blood clotted in the presence of calcium Natural inhibitors of fibrinolysis inhibit the activa- ions, but is the same as that in serum obtained in the tion of plasminogen to plasmin (inhibitor of plasmino- absence of calcium ions. gen activation or activator inhibitor) or act upon the Radiolabeled a2-plasmin inhibitor was covalently plasmin already formed (plasmin inhibitors). Although bound to fibrin only when calcium ions were present the presence of the former inhibitors in plasma (1) has at the time of clotting of plasma or fibrinogen. Whereas, not been fully substantiated, the presence of plasmin when batroxobin, a snake venom enzyme that lacks the inhibitors has been firmly established (2). ability to activate fibrin-stabilizing factor, was used for Among various proteinase inhibitors capable of in- clotting fibrinogen, the binding was not observed. When hibiting plasmin activity, two inhibitors, a2-plasmin in- fibrin-stabilizing, factor-deficient plasma was clotted, hibitor (a2PI)1 and a2-macroglobulin are considered to the specific binding of a2-plasmin inhibitor to fibrin be physiologically most important (2). Plasmin gener- did not occur even in the presence of calcium ions ated in a plasma milieu is effectively inhibited by these and the concentration of a2-plasmin inhibitor in serum two inhibitors, therefore fibrinogenolysis does not oc- was the same as that in plasma. cur under these conditions. Monodansyl cadaverine, a fluorescent substrate of When fibrin is formed, plasminogen activators and the fibrin-stabilizing factor, was incorporated into a2- plasminogen are adsorbed to fibrin and plasminogen plasmin inhibitor by activated fibrin-stabilizing factor. activation takes place on fibrin molecules to exert an All these findings indicate that a2-plasmin inhibitor effective fibrinolysis. This process of fibrinolysis is ef- is cross-linked to fibrin by activated fibrin-stabilizing ficiently inhibited by a2PI but not by a2-macroglobulin factor when blood is clotted. Analysis of a2-plasmin (2, 3). This hypothesis was substantiated by the dis- inhibitor-incorporated fibrin by sodium dodecyl sulfate covery of congenital deficiency of a2PI (4). The efficient gel electrophoresis showed that the inhibitor was inhibition of fibrinolysis by a2PI has been attributed mainly cross-linked to polymerized a-chains of cross- to the inhibition of binding of plasminogen to fibrin linked fibrin. Cross-linking of a2-plasmin inhibitor to by a2PI (2, 3) in addition to the rapid inactivation of fibrin renders fibrin clot less susceptible to fibrinolysis plasmin formed during plasminogen activation (2,5,6). by plasmin. In a brief, preliminary communication (7), we showed that a2PI was adsorbed to fibrin when plasma was clotted and suggested that the adsorption might be INTRODUCTION another mechanism of the efficient inhibition of fibrinol- The dissolution of fibrin deposits or thrombin in vivo ysis by a2PI. is achieved by plasmin formed from plasminogen by We now present evidence that a2PI is bound to plasminogen activator that has been added extrinsically fibrin by a covalent linkage formed by activated fibrin- for the therapeutic purposes (urokinase and strepto- stabilizing factor (FSF, blood coagulation factor XIII, kinase) or endogenously generated in the vascular trees plasma transglutaminase), and suggest that the covalent (blood activators or vascular activator). Regardless of binding of a2PI to fibrin contributes significantly to the the manner in which plasminogen is activated, dis- stability of fibrin. ' Abbreviations used in this paper: a2PI, a2-plasmin inhibi- Received for publication 9 July 1979 and in revised form tor; FSF, fibrin-stabilizing factor; MDC, monodansyl cadaver- I October 1979. ine; SDS, sodium dodecyl sulfate. 290 J. Clin. Invest. (D The American Society for Clinical Investigation, Inc. 0021-9738/80/02/0290/08 $1.00 Volume 65 February 1980 290-297 METHODS 50 U/ml, CaCl2, 0.375 M). FSF-deficient plasma was kindly supplied by Dr. M. Fujimaki, Department of Clinical Pa- Fibrinogen. Human fraction 1-4 prepared by the method thology, Tokyo Medical College. The blood was obtained from of Blomback and Blomback (8) or human Cohn's fraction I a patient with congenital deficiency of FSF and immediately (Green Cross Corp., Osaka) was used as fibrinogen preparation mixed with 1/10 vol of 3.8% sodium citrate to obtain citrated after removing contaminating plasminogen from the prepara- plasma. When the fibrin formed by the addition of thrombin tion using lysine-Sepharose (Pharmacia Fine Chemicals, Inc., and calcium ions to this plasma was analyzed by sodium do- Piscataway, N. J.) (9). The concentration of FSF present in decyl sulfate gel electrophoresis (14), no formation of y-chain the solution of 250 mg/100 ml of fraction 1-4 or Cohn's fraction dimer of fibrin was observed, indicating the absence of FSF I was 50 or 80% of normal standard plasma, respectively, when in the patient's plasma. Serum was obtained by incubating assayed by an antibody neutralization method (10) using a 1 ml of plasma at 37°C for 2 h with 25 ,ul of 0.5 M CaCl2. Clotting Factor XIII-test kit supplied by Behringwerke AG, a2PI-deficient plasma was obtained from a patient with con- Marburg. Cold-insoluble globulin was not detected in the genital deficiency of a2PI (4). Aprotinin (Trasylol, Bayer fraction 1-4 preparation by double immunodiffusion, single Chemical, Tokyo) was added to the plasma in a ratio of 5 IAI radial immunodiffusion or counter immunoelectrophoresis aprotinin (5,000 U/ml):1 ml plasma to avoid spontaneous using specific antiserum against cold-insoluble globulin (11). fibrinolysis when the plasma was clotted. There was also no characteristic band of subunit cold-in- Radioiodination of protein. Purified a2PI and fibrinogen soluble globulin with mol wt 215,000 when the preparation (fraction 1-4) were radioiodinated by the method of Thorell was analyzed by sodium dodecyl sulfate gel electrophoresis and Johansson (17) or the solid-state lactoperoxidase method with reduction (11). Cohn's fraction I was clearly con- of David (18), respectively, using lactoperoxidase (Calbiochem- taminated with cold-insoluble globulin. Fibrinogen was Behring Corp., American Hoechst Corp., San Diego, Calif.) dissolved in barbital-buffered saline (0.005 M barbital-acetic and 125I-Na (17 Ci/ml) (New England Nuclear, Boston, Mass.). acid-0. 14 M NaCl, pH 7.4) and the concentration was adjusted The labeled a2PI preparation had the radioactivity of 1.1 to 1.5% (wt/vol) clottable protein. x 107 cpm/,ug. Specific activity of a2PI before and after radio- Thrombin. Purified thrombin was prepared from bovine iodination was 1136 U/A280 and 1,050 U/A2s0, respectively. thrombin preparation (Parke, Davis & Co., Detroit, Mich.) a2PI concentration of the labeled a2PI preparation was 60 according to the method of Lundblad (12), and was stored ixg/ml. The labeled fibrinogen preparation had a radioactivity as a 200 U/ml solution in 50% glycerol at -200C. of 6.3 x 104 cpm/Ig. Clottability (97%) was not changed, and Plasmin. Plasminogen was purified from human whole fibrinogen concentration of the labeled preparation was 316 plasma by affinity chromatography using lysine-coupled mg/100 ml clottable protein. Sepharose 4B (13), and was activated with urokinase-coupled Measurement of the binding of a2PI tofibrin. When plasma Sepharose according to the described method (5). Total con- was used, 1 ml of plasma was mixed in a counting vial for version of plasminogen to two-chain plasmin form was as- radioactivity with 5 Al of radiolabeled a2PI and the mixture certained by sodium dodecyl sulfate gel electrophoresis with was clotted by the addition of 40 Iul of thrombin (50 U/ml) reduction (14). Plasmin thus obtained was stored at -20°C as a or a thrombin-calcium mixture (thrombin 50 U/ml, CaCl2 0.375 65 caseinolytic U/ml solution (15) in buffered saline contain- M).
Recommended publications
  • (12) United States Patent (10) Patent No.: US 6,395,889 B1 Robison (45) Date of Patent: May 28, 2002
    USOO6395889B1 (12) United States Patent (10) Patent No.: US 6,395,889 B1 Robison (45) Date of Patent: May 28, 2002 (54) NUCLEIC ACID MOLECULES ENCODING WO WO-98/56804 A1 * 12/1998 ........... CO7H/21/02 HUMAN PROTEASE HOMOLOGS WO WO-99/0785.0 A1 * 2/1999 ... C12N/15/12 WO WO-99/37660 A1 * 7/1999 ........... CO7H/21/04 (75) Inventor: fish E. Robison, Wilmington, MA OTHER PUBLICATIONS Vazquez, F., et al., 1999, “METH-1, a human ortholog of (73) Assignee: Millennium Pharmaceuticals, Inc., ADAMTS-1, and METH-2 are members of a new family of Cambridge, MA (US) proteins with angio-inhibitory activity', The Journal of c: - 0 Biological Chemistry, vol. 274, No. 33, pp. 23349–23357.* (*) Notice: Subject to any disclaimer, the term of this Descriptors of Protease Classes in Prosite and Pfam Data patent is extended or adjusted under 35 bases. U.S.C. 154(b) by 0 days. * cited by examiner (21) Appl. No.: 09/392, 184 Primary Examiner Ponnathapu Achutamurthy (22) Filed: Sep. 9, 1999 ASSistant Examiner William W. Moore (51) Int. Cl." C12N 15/57; C12N 15/12; (74) Attorney, Agent, or Firm-Alston & Bird LLP C12N 9/64; C12N 15/79 (57) ABSTRACT (52) U.S. Cl. .................... 536/23.2; 536/23.5; 435/69.1; 435/252.3; 435/320.1 The invention relates to polynucleotides encoding newly (58) Field of Search ............................... 536,232,235. identified protease homologs. The invention also relates to 435/6, 226, 69.1, 252.3 the proteases. The invention further relates to methods using s s s/ - - -us the protease polypeptides and polynucleotides as a target for (56) References Cited diagnosis and treatment in protease-mediated disorders.
    [Show full text]
  • United States Patent (19) 11 Patent Number: 6,019,993 Bal (45) Date of Patent: Feb
    USOO6O19993A United States Patent (19) 11 Patent Number: 6,019,993 Bal (45) Date of Patent: Feb. 1, 2000 54) VIRUS-INACTIVATED 2-COMPONENT 56) References Cited FIBRIN GLUE FOREIGN PATENT DOCUMENTS 75 Inventor: Frederic Bal, Vienna, Austria O 116 263 1/1986 European Pat. Off.. O 339 607 4/1989 European Pat. Off.. 73 Assignee: Omrix Biopharmaceuticals S.A., O 534 178 3/1993 European Pat. Off.. Brussels, Belgium 2 201993 1/1972 Germany. 2 102 811 2/1983 United Kingdom. 21 Appl. No.: 08/530,167 PCTUS85O1695 9/1985 WIPO. PCTCH920.0036 2/1992 WIPO. 22 PCT Filed: Mar. 27, 1994 PCT/SE92/ 00441 6/1992 WIPO. 86 PCT No.: PCT/EP94/00966 PCTEP9301797 7/1993 WIPO. S371 Date: Nov.30, 1995 Primary Examiner-Carlos Azpuru Attorney, Agent, or Firm-Jacobson, Price, Holman, & S 102(e) Date: Nov.30, 1995 Stern, PLLC 87 PCT Pub. No.: WO94/22503 57 ABSTRACT PCT Pub. Date: Oct. 13, 1994 A two-component fibrin glue for human application includes 30 Foreign Application Priority Data a) a component A containing i) a virus-inactivated and concentrated cryoprecipitate that contains fibrinogen, and ii) Mar. 30, 1993 EP European Pat. Off. .............. 93105298 traneXamic acid or a pharmaceutically acceptable Salt, 51 Int. Cl." A61F 2/06; A61K 35/14 thereof, and b) component B containing a proteolytic 52 U s C - - - - - - - - - - - - - - - - - - - - - - - - - - -424/426. 530,380, 530/381: enzyme that, upon combination with component A, cleaves, O X O -- O - - - s 530(383. 530ass Specifically, fibrinogen present in the cryoprecipitate of 58) Field of Search 424/426,530,380 component A, thereby, effecting a fibrin polymer.
    [Show full text]
  • Safety and Efficacy of Prothrombin Complex Concentrate As First-Line
    Cappabianca et al. Critical Care (2016) 20:5 DOI 10.1186/s13054-015-1172-6 RESEARCH Open Access Safety and efficacy of prothrombin complex concentrate as first-line treatment in bleeding after cardiac surgery Giangiuseppe Cappabianca1†, Giovanni Mariscalco2*† , Fausto Biancari3, Daniele Maselli4, Francesca Papesso1, Marzia Cottini1, Sandro Crosta5, Simona Banescu5, Aamer B. Ahmed6 and Cesare Beghi1 Abstract Background: Bleeding after cardiac surgery requiring surgical reexploration and blood component transfusion is associated with increased morbidity and mortality. Although prothrombin complex concentrate (PCC) has been used satisfactorily in bleeding disorders, studies on its efficacy and safety after cardiopulmonary bypass are limited. Methods: Between January 2005 and December 2013, 3454 consecutive cardiac surgery patients were included in an observational study aimed at investigating the efficacy and safety of PCC as first-line coagulopathy treatment as a replacement for fresh frozen plasma (FFP). Starting in January 2012, PCC was introduced as solely first-line treatment for bleeding following cardiac surgery. Results: After one-to-one propensity score–matched analysis, 225 pairs of patients receiving PCC (median dose 1500 IU) and FFP (median dose 2 U) were included. The use of PCC was associated with significantly decreased 24-h post-operative blood loss (836 ± 1226 vs. 935 ± 583 ml, p < 0.0001). Propensity score–adjusted multivariate analysis showed that PCC was associated with significantly lower risk of red blood cell (RBC) transfusions (odds ratio [OR] 0.50; 95 % confidence interval [CI] 0.31–0.80), decreased amount of RBC units (β unstandardised coefficient −1.42, 95 % CI −2.06 to −0.77) and decreased risk of transfusion of more than 2 RBC units (OR 0.53, 95 % CI 0.38–0.73).
    [Show full text]
  • Serine Proteases with Altered Sensitivity to Activity-Modulating
    (19) & (11) EP 2 045 321 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 08.04.2009 Bulletin 2009/15 C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12Q 1/37 (2006.01) (21) Application number: 09150549.5 (22) Date of filing: 26.05.2006 (84) Designated Contracting States: • Haupts, Ulrich AT BE BG CH CY CZ DE DK EE ES FI FR GB GR 51519 Odenthal (DE) HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI • Coco, Wayne SK TR 50737 Köln (DE) •Tebbe, Jan (30) Priority: 27.05.2005 EP 05104543 50733 Köln (DE) • Votsmeier, Christian (62) Document number(s) of the earlier application(s) in 50259 Pulheim (DE) accordance with Art. 76 EPC: • Scheidig, Andreas 06763303.2 / 1 883 696 50823 Köln (DE) (71) Applicant: Direvo Biotech AG (74) Representative: von Kreisler Selting Werner 50829 Köln (DE) Patentanwälte P.O. Box 10 22 41 (72) Inventors: 50462 Köln (DE) • Koltermann, André 82057 Icking (DE) Remarks: • Kettling, Ulrich This application was filed on 14-01-2009 as a 81477 München (DE) divisional application to the application mentioned under INID code 62. (54) Serine proteases with altered sensitivity to activity-modulating substances (57) The present invention provides variants of ser- screening of the library in the presence of one or several ine proteases of the S1 class with altered sensitivity to activity-modulating substances, selection of variants with one or more activity-modulating substances. A method altered sensitivity to one or several activity-modulating for the generation of such proteases is disclosed, com- substances and isolation of those polynucleotide se- prising the provision of a protease library encoding poly- quences that encode for the selected variants.
    [Show full text]
  • Correlation of the Production of Plasminogen Activator with Tumor Metastasis in Bi 6 Mouse Melanoma Cell Lines
    [CANCER RESEARCH 40, 288-292, February 1980 0008-5472/80/0040-0000$02.00 Correlation of the Production of Plasminogen Activator with Tumor Metastasis in Bi 6 Mouse Melanoma Cell Lines Bosco Shang Wang,2 Gerard A. McLoughlin,3 Jerome P. Richie, and John A. Mannick Deportment of Surgery, Peter Bent Brigham Hospital (B. S. W., G. A. M., J. P. R., J. A. M.j, and Department of Pathology (B. S. W.j, Harvard Medical School, Boston, Massachusetts 02115 ABSTRACT circulating cancer cells were easily detected in patients whose blood had a higher level of FA. The correlation between the production of plasminogen ac Although the FA of a tumor has been suggested as correlat tivator (PA) of tumors and their metastatic potential was stud ing with its metastatic potential, direct and quantitative evi ied. Bi 6 melanoma cells and ‘‘Bi6 mets' ‘cells (harvested from dence to support this hypothesis is lacking. Therefore, we have the pulmonary metastatic nodules of C57BL/6J mice bearing investigated this issue by comparing the FA of the Bi 6 mouse Bi 6 isografts) were examined with respect to their fibrinolytic melanoma and several of its sublines varying in their potential activity (FA) in tissue culture. Bi 6 mets cells had a significantly for forming metastases. higher FA than did Bi 6 cells. Fl (a Bi 6 subline with a lower incidence of metastasis) and Fl 0 (a highly metastatic Bi 6 subline) were also studied. Fl 0 cells produced more FA than MATERIALS AND METHODS did Fi cells. The difference between the FA's of these tumors Tumors.
    [Show full text]
  • A Study on Ulinastatin in Preventing Post ERCP Pancreatitis
    International Journal of Advances in Medicine Vedamanickam R et al. Int J Adv Med. 2017 Dec;4(6):1528-1531 http://www.ijmedicine.com pISSN 2349-3925 | eISSN 2349-3933 DOI: http://dx.doi.org/10.18203/2349-3933.ijam20175083 Original Research Article A study on ulinastatin in preventing post ERCP pancreatitis R. Vedamanickam1, Vinoth Kumar2*, Hariprasad2 1Department of Medicine, 2Department of Gasto and Hepatology , SREE Balaji Medical College and Hospital, Chrompet, Chennai, Tamil Nadu, India Received: 19 September 2017 Accepted: 25 October 2017 *Correspondence: Dr. Vinothkumar, E-mail: [email protected] Copyright: © the author(s), publisher and licensee Medip Academy. This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT Background: Pancreatitis remains the major complication of endoscopic retrograde cholangiopancreatography (ERCP), and hyperenzymemia after ERCP is common. Ulinastatin, a protease inhibitor, has proved effective in the treatment of acute pancreatitis. The aim of this study was to assess the efficacy of ulinastatin, compare to placebo study to assess the incidence of complication due to ERCPP procedure. Methods: In this study a randomized placebo controlled trial, patients undergoing the first ERCP was randomizing to receive ulinastatin one lakh units (or) placebo by intravenous infusion one hour before ERCP for ten minutes duration. Clinical evaluation, serum amylase, ware analysed before the procedure 4 hours and 24 hours after the procedure. Results: Total of 46 patients were enrolled (23 in ulinastatin and 23 in placebo group).
    [Show full text]
  • Effect of Intravenous Thrombolysis with Alteplase on Clinical Efficacy
    Brazilian Journal of Medical and Biological Research (2021) 54(5): e10000, http://dx.doi.org/10.1590/1414-431X202010000 ISSN 1414-431X Research Article 1/8 Effect of intravenous thrombolysis with alteplase on clinical efficacy, inflammatory factors, and neurological function in patients with acute cerebral infarction Jinhua Wang1 00 00, Xia Fang2 00 , Dongliang Wang1 00 , and Yuan Xiao1 00 1Department of Neurology, The People’s Hospital of Beilun District, Beilun Branch Hospital of The First Affiliated Hospital, Zhejiang University School of Medicine, Ningbo, Zhejiang Province, China 2Department of Gynecology, The People’s Hospital of Beilun District, Beilun Branch Hospital of The First Affiliated Hospital, Zhejiang University School of Medicine, Ningbo, Zhejiang Province, China Abstract This study aimed to explore the effect of intravenous thrombolysis with alteplase on clinical efficacy, inflammatory factors, and neurological function in patients with acute cerebral infarction. A total of 120 patients with acute cerebral infarction were divided into two groups by the random number table method, with 60 patients in each group: observation group (intravenous thrombolysis with alteplase) and control group (intravenous thrombolysis with batroxobin). The clinical efficacy after a 14-day treatment was observed. Serum C-reactive protein (CRP), tumor necrosis factor a (TNF-a), interleukin-6 (IL-6), CD62p, GMP- 140, and neuron-specific enolase (NSE) were measured. Scores of National Institutes of Health Stroke Scale (NIHSS), Mini- Mental State Examination (MMSE), and Montreal Cognitive Assessment (MoCA) were determined. The total effective rate in the observation group was 81.67%, which was higher than the 61.67% in the control group (Po0.05).
    [Show full text]
  • Distinctive Role of Histidine-16 of the B,8 Chain of Fibrinogen in the End-To
    Proc. Nati. Acad. Sci. USA Vol. 83, pp. 591-593, February 1986 Biochemistry Distinctive role of histidine-16 of the B,8 chain of fibrinogen in the end-to-end association of fibrin (polymerization/chemical modification/affinity chromatography) AKIRA SHIMIZU, YUJI SAITO, AND YUJI INADA Laboratory of Biological Chemistry, Tokyo Institute of Technology, Ookayama, Meguro-ku, Tokyo 152, Japan Communicated by John D. Ferry, September 12, 1985 ABSTRACT Photooxidation of fibrinogen reduced the includes 17th to 19th amino acid residues in Aa chain of batroxobin-induced fibrin polymerization. The fibrin fragment fibrinogen, specifically inhibits fibrin association and has des-AB N-DSK, which contains the binding sites termed A and proved itselfuseful for the elucidation ofassociation sites (7). B, lost the ability to bind to the site termed a in fibrinogen- These examples stress the critical role played by arginine-19 Sepharose upon the oxidation of histidine-16 in the BP chain of of Aa chain and some residues adjacent to it in the NH2- fibrinogen [Shimizu, A., Saito, Y., Matsushima, A. & Inada, terminal domain. On the other hand, very short fragments Y. (1983) J. Biol. Chem. 258, 7915-7917]. Some of the derived from the y chain of the COOH-terminal domain fragments, which became unable to bind to fibrinogen-Seph- (threonine-374 to valine-411 or threonine-374 to glutamic arose due to the destruction of site A, however, retained the acid-396) quite efficiently inhibited the association and sug- ability to bind to D-dimer-Sepharose, which contains both sites gested the importance ofthat region for the association (8, 9).
    [Show full text]
  • Antiproteases in Preventing Post-ERCP Acute Pancreatitis
    JOP. J Pancreas (Online) 2007; 8(4 Suppl.):509-517. ROUND TABLE Antiproteases in Preventing Post-ERCP Acute Pancreatitis Takeshi Tsujino, Takao Kawabe, Masao Omata Department of Gastroenterology, Faculty of Medicine, University of Tokyo. Tokyo, Japan Summary there is no other randomized, placebo- controlled trial on ulinastatin under way. Pancreatitis remains the most common and Large scale randomized controlled trials potentially fatal complication following revealed that both the long-term infusion of ERCP. Various pharmacological agents have gabexate and the short-term administration of been used in an attempt to prevent post-ERCP ulinastatin may reduce pancreatic injury, but pancreatitis, but most randomized controlled these studies involve patients at average risk trials have failed to demonstrate their of developing post-ERCP pancreatitis. efficacy. Antiproteases, which have been Additional research is needed to confirm the clinically used to manage acute pancreatitis, preventive efficacy of these antiproteases in would theoretically reduce pancreatic injury patients at a high risk of developing post- after ERCP because activation of proteolytic ERCP pancreatitis. enzymes is considered to play an important role in the pathogenesis of post-ERCP pancreatitis. Gabexate and ulinastatin have Introduction recently been evaluated regarding their efficacy in preventing post-ERCP ERCP is widely performed for the diagnosis pancreatitis. Long-term (12 hours) infusion of and management of various pancreaticobiliary gabexate significantly decreased the incidence diseases. Early complications after ERCP of post-ERCP pancreatitis; however, no include acute pancreatitis, bleeding, prophylactic effect was observed for short- perforation, and infection (cholangitis and term infusion (2.5 and 6.5 hours). These cholecystitis) [1, 2]. Of these ERCP-related results may be due to the short-life of complications, pancreatitis remains the most gabexate (55 seconds).
    [Show full text]
  • Exogenous Procoagulant Factors As Therapeutic and Biotechnological
    isord D ers od & lo T r B f a n o s l f a u n s r Journal of i o u n o Chudzinski-Tavassi et al., J Blood Disorders Transf 2014, 5:5 J ISSN: 2155-9864 Blood Disorders & Transfusion DOI: 10.4172/2155-9864.1000209 Review Article OpenOpen Access Access Exogenous Procoagulant Factors as Therapeutic and Biotechnological Tools Ana Marisa Chudzinski-Tavassi*, Linda Christian Carrijo Carvalho, Miryam Paola Alvarez-Flores and Sonia Aparecida de Andrade Laboratory of Biochemistry and Biophysics, Butantan Institute, São Paulo, Brazil Abstract A diversity of animal venoms and secretions has been described to affect the hemostatic system with actions on blood coagulation and fibrinolytic pathways. These biological materials are rich sources of proteins and peptides with distinct biochemical properties, which have a biological function for the animal. Snake venoms are one of the richest sources of exogenous hemostatic factors, especially procoagulant proteins. Insects are another important source of proteins and peptides targeting the hemostatic system. Exogenous procoagulant factors have a large functional diversity and present potential applications in health and biotechnology. They have been important tools for the diagnosis and therapy of several blood coagulation disorders. Recently, many studies have pointed out that exogenous hemostatic factors can also display non-hemostatic functions, bringing new perspectives for the study of these molecules. Keywords: Exogenous hemostatic factors; Coagulation; Fibrinolysis; venom production, storage and
    [Show full text]
  • Mechanism of Batroxobin Binding to Fibrinogen and Fibrin 1
    JBC Papers in Press. Published on April 23, 2013 as Manuscript M113.464750 The latest version is at http://www.jbc.org/cgi/doi/10.1074/jbc.M113.464750 Mechanism of batroxobin binding to fibrinogen and fibrin BATROXOBIN BINDS FIBRIN WITH HIGHER AFFINITY AND PROMOTES CLOT EXPANSION TO A GREATER EXTENT THAN THROMBIN Trang T. Vu1,2, Alan R. Stafford1,3, Beverly A. Leslie1,3, Paul Y. Kim1,3, James C. Fredenburgh1,3 and Jeffrey I. Weitz1,2,3,4† Thrombosis and Atherosclerosis Research Institute1 and the Departments of Medical Sciences,2 Medicine,3 and Biochemistry and Biomedical Sciences,4 McMaster University, Hamilton, Ontario, Canada. Running title: Mechanism of batroxobin binding to fibrinogen and fibrin To whom correspondence should be addressed: Dr. Jeffrey Weitz, Thrombosis and Atherosclerosis Research Institute, 237 Barton St. E., Hamilton, Ontario L8L 2X2, Canada Phone: 905-574-8550 Fax: (905) 575-2646 E-mail: [email protected] Downloaded from Keywords: Batroxobin, thrombin, fibrin(ogen), fibrinopeptides Background: Snake venom protease γA/γ'-fibrin(ogen) than γA/γA-fibrin(ogen). batroxobin clots fibrinogen in a manner In contrast, batroxobin binds both http://www.jbc.org/ distinct from thrombin. fibrin(ogen) isoforms with similar high Results: Batroxobin binds fibrin(ogen) with affinity (Kd values of about 0.5 μM) even higher affinity than thrombin and promotes though it does not interact with the γ'-chain. greater clot expansion. The batroxobin binding sites on Conclusion: Batroxobin’s distinctive fibrin(ogen) only partially overlap with by guest on July 22, 2019 interaction with fibrin(ogen) may contribute to those of thrombin because thrombin its unique pattern of fibrinopeptide release.
    [Show full text]
  • EXPERT COMMITTEE on BIOLOGICAL STANDARDIZATION Geneva, 17 to 21 October 2016
    WHO/BS/2016.2282 ENGLISH ONLY EXPERT COMMITTEE ON BIOLOGICAL STANDARDIZATION Geneva, 17 to 21 October 2016 An international collaborative study to calibrate the WHO 2nd International Standard for Ancrod (15/106) and the WHO Reference Reagent for Batroxobin (15/140) Craig Thelwell1ᶲ, Colin Longstaff1ᶿ, Peter Rigsby2, Matthew Locke1 and Sally Bevan1 1Biotherapeutics Group, Haemostasis Section and 2Biostatistics Section, National Institute for Biological Standards and Control, South Mimms, Herts EN6 3QG, UK ᶲProject leader for Ancrod; ᶿProject leader for Batroxobin NOTE: This document has been prepared for the purpose of inviting comments and suggestions on the proposals contained therein, which will then be considered by the Expert Committee on Biological Standardization (ECBS). Comments MUST be received by 16 September 2016 and should be addressed to the World Health Organization, 1211 Geneva 27, Switzerland, attention: Technologies, Standards and Norms (TSN). Comments may also be submitted electronically to the Responsible Officer: Dr C M Nübling at email: [email protected] © World Health Organization 2016 All rights reserved. Publications of the World Health Organization are available on the WHO web site (www.who.int) or can be purchased from WHO Press, World Health Organization, 20 Avenue Appia, 1211 Geneva 27, Switzerland (tel.: +41 22 791 3264; fax: +41 22 791 4857; e-mail: [email protected]). Requests for permission to reproduce or translate WHO publications – whether for sale or for noncommercial distribution – should be addressed to WHO Press through the WHO web site: (http://www.who.int/about/licensing/copyright_form/en/index.html). The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries.
    [Show full text]