DOCSLIB.ORG

Studies on Interaction Between Coagulation Factor X and Plasmin*)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Studies on Interaction Between Coagulation Factor X and Plasmin*) Hiroshima Journal of Medical Sciences 785 Vol. 33, No. 4, 785,......,788, December, 1984 HIJM 33-113 Studies on Interaction between Coagulation Factor X and Plasmin*) Sumiyoshi TAKASUGI, Masayuki NISHIKI, Motoo KANAO, Tsuneo OKUMICHI and Haruo EZAKI The 2nd Department of Surgery, Hiroshima University School of Medicine, 1-2-3, Kasumi, Minami-ku, Hiroshima 734, Japan (Received September 25, 1984) Key words: Factor X, Plasmin, Factor Xa ABSTRACT Coagulation factor X was isolated from normal human plasma by a two-step sequential affinity column chromatography method. The concentration of factor X was confirmed to be 115 units per litter of plasma. In the present study, it was first confirmed that factor X could be activated to factor Xa by Lys-plasmin. by the methods previously described12>. Lys­ INTRODUCTION plasmin (specific activity; 30 IU /mg protein) Coagulation factor X circulate in plasma in was a gift from Professor Kenneth C, Robbins an inactive or zymogen form, and must be (Michael Reese Research Foundation, Chicago, converted to an active form, factor Xa, by com­ USA). ponents of the intrinsic or extrinsic pathway. Preparation of plasma: Factor Xa, in turn, hydrolyzes prothrombin to Venous blood samples were drawn from nor­ thrombin in the presence of calcium ions, phos­ mal persons, volunteers and a 52-years-old man pholipid, and a co-factor, factor V. Thrombin who was diagnosed as acute pancreatitis fol­ then converts fibrinogen to fibrin, which be­ lowed by disseminated intravascular coagulation comes insoluble and precipitates in plasma to (DIC). One volume of 3. 8% sodium citrate form a clot. The activation of factor X by was added to nine volumes of blood sample, means of the intrinsic pathway is catalyzed by and plasma was obtained by centrifugation of factor IXa and VIII in the presence of phos­ the mixture at 1580 g for 10 min at 4°C. Plas­ pholipid and calcium ions4• 6>, and the activation tic equipment and previously siliconized glas­ by extrinsic pathway is catalyzed by factors sware and syringe needles were used to prevent VII and tissue factor in the presence of phos­ the activation of coagulation or fibrinolysis 1 pholipid and calcium ions • 4>. On the other factors. hand, it has been reported that the coagulant Determination of amidolytic activity by factor protein of the venom from the Russel's viper7> Xa: and trypsin4> also capable of activating factor Factor Xa amidolytic activity was determined X. This paper describes about the activation by a modification of the endpoint method de­ of factor X by plasmin. scribed by Claeson et al. 2> with Bz-Ile-Glu­ Gly-Arg-pNA, measuring released p-nitroani­ MATERIALS AND METHODS line (pNA). The system contained 0. 75 ml of Reagents: 0. 05 M Tris-HCl buffer, pH 7. 5 containing 0. 15 Pyro-Glu-Gly-Arg-pNA (Kabi, S-2444), H­ M NaCl and 10 KIU/ml of aprotinin, 0. 1 ml D-Phe-L--Pip-L-Arg-pNA (Kabi, S-2238), Bz­ of sample solution, 0. 05 ml of 0. 1% gelatine Ile-Glu-Gly-Arg-pNA (Kabi, S-2222), Aprotin­ solution containing 10 KIU/ml of aprotinin and in (Bayer), Zinc-Chelate-Agarose (Pierce) were 0. 1 ml of 3 mM S-2222 solution; after incuba­ purchased. ACH-Sepharose 4B was prepared tion for 5 min at 37° C, 0. 1 ml of 50 % acetic 786 S. Takasugi et al. acid was added to stop the reaction; the absorb­ original plasma. To confirm the presence of ance was read at 405 nm, S-2222 is a substrate UK-zymogen in the adsorbed and eluted frac­ for factor Xa, is cleaved, by factor Xa ten times tions, the changes of S-2444 amidolytic activity as rapidly as by thronibin, and six times as in ·Zinc-chelate adsorbed and· eluted fractions rapidly as by a VIIa1 3>. was about, 4-5 times increased after incubation Factor X, a zymogen form of factor Xa, was with 3 IU of Lys-plasmin for 15 min at 37° C. measured by enhancement of S-2222 amidolytic The authors considered that the increase of activity after activation of factor X to factor the S-2444 amidolytic activity might be a phe­ Xa by Lys-plasmin treatment. Treatment with nomenon induced by the activation of UK­ Lys-plasmin was carried out by mixing 0.1 ml zymogen to active UK, because S-2444 is useful of the sample solution and 0. 1 ml of Lys-plas­ for determining UK activity, and plasmin has min (0. 1 mg/ml, 3 IU/ml), and incubating at been confirmed to activate the UK-zymogen to 37° C for 15 min. The reaction was stopped by active UK11 ' 15>. To further confirm about this, adding 100 KIU of aprotinin to inhibit plasmin. the authors applied the Zinc-chelate adsorbed S-2444 and S-2238 amidolytic activities were and eluted fractions on a monospecific anti­ determined by quite the same method as S- UK-IgG-Sepharose 4B column. Unexpected­ 2222. ly, UK-zymogen-like protein in adsorbed and eluted fractions failed in binding to anti-UK­ RESULTS AND DISCUSSIONS IgG-Sepharose 4B column, indicating that this Isolation of factor X from normal human protein is not UK-zymogen, because UK-zymo­ plasma, and activation of factor X to Xa by gen possess the same antigenicity with active Lys-plasmin: UK11,1s>. : Zinc-chelate-Sepharose 4B affinity chromato­ It is well known that S-2444 can be hydro­ 9 graphy was introduced by Porath et aL > and lyzed not only by UK, but also by plasmin, has been used as the isolation method of tissue­ kallikrein, thrombin and factor Xa. Since plasma type plasminogen activator10>. In the present plasmin and kallikrein can be inhibited by aprotin­ study, the authors first tried to isolate urokinase in, there remains the possibility that UK-zymo­ 11 (UK)-zymogen 115) from normal human plasma gen-like protein may be clotting factors, such as using this method as follows; 20 ml of citrated thrombin and factor Xa. Therefore, the authors human plasma was passed through the Zinc­ examined whether this protein is thrombin or chelate-Sepharose 4B column (2 x 5 cm) equili­ factor Xa using their specific substrates, S-2238 brated with 0. 02 M Tris-HCl buffer, pH 7. 5 and S-2222, respectively. As shown in Table containing 1 M NaCL After washing with ex­ 1, UK-zymogen-like enzyme did not hydrolyze cess amounts of the equilibration buffer until S-2238, but S-2222, indicating that this protein absorbance at 280 nm was zero, adsorbed frac­ might be factor X or Xa. In the present study, tions were eluted with the same equilibration the authors further purified factor X from Zinc­ buffer containing 0. 05 M Imidazole. Adsorbed chelate adsorbed and eluted fractions using and eluted factions were collected and dialyzed ACH-Sepharose 4B as follows; 30 ml of Zinc­ against .the phosphate buffer saline (PBS) at chelate adsorbed and eluted fractions was passed 4° C overnight. By this affinity chromatography through the ACH-Sepharose 4B column (2 x 5 method, about 1. 7 mg of protein was isolated cm) equilibrated with 0. 1 M phosphate buffer, in adsorbed and eluted fractions from 20 ml of pH 7. 4 containing 2 M NaCL Sixty ml of un- Table 1. Amidolytic activity of Zinc-chelate adsorbed-eluted fraction S-2444 S-2238 S-2222 amidolytic activity amidolytic activity amidolytic activity LIA405/5 min/O. l * . before Lys-plasmin treatment 0.011 0.007 0.053 after Lys-plasmin treatment 0.050 0.006 0.256 * 0. l ml of sample solution is proportionate to 2 ml of orginal plasma Coagulation Factor X and Plasmin 787 Table 2. Purification of factor X from normal human plasma by a two-step sequential affinity chromatography Zinc-chelate-Sepharose 4B ACH-Sepharose 4B unadsorbed adsorbed-eluted unadsorbed adsorbed-eluted fractions fractions fractions fractions total protein (mg) 1360 1.7 1.1 0.6 S-2222 amidolytic activity (U)** ND* 2.6 2.3 0 Data is average of 11 purification experiments. Total volume of original plasma sample is 20 ml. *ND ; not determined, **U; l unit was defined as the amount of enzyme which produces L1 A405 /5 min= 1. 0 after Lys-plasmin treatment under standard condition described in Meterials and Methods. adsorbed fractions were collected with equilibra­ though the activation mechanism of factor X tion buffer. After washing with equilibration to factor Xa is still unclear, this phenomenon buffer until absorbance at 280 nm was zero, makes us speculate the possibility that plasmin adsorbed fractions were eluted with the equili­ may play an important role in maintenance of bration buffer containing 8 M urea and 30 ml the hemostatic balance as the factor of feed fractions were collected. Both unadsorbed and back from fibrinolysis to coagulation. adsorbed and eluted fractions were dialyzed Isolation of factor X from patient plasma against PBS (2L x 2) overnight at 4° C. The with acute pancreatitis followed by DIC: precipitates formed during dialysis were re­ The authors tried to isolate factor X from moved out by centrifugation 10, 000 rpm for 30 patient plasma with acute pancreatitis followed min and concentrated to 3 ml by Carbowax by DIC, and confirmed that no factor X-like 20, 000. Table 2 summarizes the purification clotting factors could be isolated from patient of factor X by this two-step sequential affinity plasma by a two-step sequential affinity chro­ chromatography method. As shown in Table matography. Serial studies of the level of factor 2, factor X could not bind to ACH-Sepharose X in DIC show significant changes in activity, 4B, and 2, 3 units of factor X was isolated but in view of the many circumstances that from 20 ml of original plasma.
Load more

Recommended publications
  • The Central Role of Fibrinolytic Response in COVID-19—A Hematologist’S Perspective
    International Journal of Molecular Sciences Review The Central Role of Fibrinolytic Response in COVID-19—A Hematologist’s Perspective Hau C. Kwaan 1,* and Paul F. Lindholm 2 1 Division of Hematology/Oncology, Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA 2 Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA; [email protected] * Correspondence: [email protected] Abstract: The novel coronavirus disease (COVID-19) has many characteristics common to those in two other coronavirus acute respiratory diseases, severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). They are all highly contagious and have severe pulmonary complications. Clinically, patients with COVID-19 run a rapidly progressive course of an acute respiratory tract infection with fever, sore throat, cough, headache and fatigue, complicated by severe pneumonia often leading to acute respiratory distress syndrome (ARDS). The infection also involves other organs throughout the body. In all three viral illnesses, the fibrinolytic system plays an active role in each phase of the pathogenesis. During transmission, the renin-aldosterone- angiotensin-system (RAAS) is involved with the spike protein of SARS-CoV-2, attaching to its natural receptor angiotensin-converting enzyme 2 (ACE 2) in host cells. Both tissue plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) are closely linked to the RAAS. In lesions in the lung, kidney and other organs, the two plasminogen activators urokinase-type plasminogen activator (uPA) and tissue plasminogen activator (tPA), along with their inhibitor, plasminogen activator 1 (PAI-1), are involved. The altered fibrinolytic balance enables the development of a hypercoagulable Citation: Kwaan, H.C.; Lindholm, state.
    [Show full text]
  • Assembly of an Integrated Human Lung Cell Atlas Reveals That
    medRxiv preprint doi: https://doi.org/10.1101/2020.06.02.20120634; this version posted June 4, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . Assembly of an integrated human lung cell atlas reveals that SARS-CoV-2 receptor is co-expressed with key elements of the kinin-kallikrein, renin-angiotensin and coagulation systems in alveolar cells Davi Sidarta-Oliveira1,2, Carlos Poblete Jara1,3, Adriano J. Ferruzzi4, Munir S. Skaf4, William H. Velander5, Eliana P. Araujo1,3, Licio A. Velloso1 1Laboratory of Cell Signaling, Obesity and Comorbidities Research Center, University of Campinas, Brazil 2 Physician-Scientist Graduate Program, School of Medical Sciences, University of Campinas, Brazil 3Nursing School, University of Campinas, Brazil 4Institute of Chemistry and Center for Computing in Engineering and Sciences University of Campinas, Brazil 5Department of Chemical and Biomolecular Engineering, University of Nebraska, Lincoln, USA Correspondence: Licio A. Velloso Laboratory of Cell Signaling, Obesity and Comorbidities Research Center, University of Campinas, Campinas, Brazil Address: Rua Carl Von Lineaus s/n, Instituto de Biologia - Bloco Z. Campus Universitário Zeferino Vaz - Barão Geraldo, Campinas - SP, 13083-864 Phone: +55 19 3521-0025 E-mail: [email protected] Abstract SARS-CoV-2, the pathogenic agent of COVID-19, employs angiotensin converting enzyme-2 (ACE2) as its cell entry receptor. Clinical data reveal that in severe COVID- 19, SARS-CoV-2 infects the lung, leading to a frequently lethal triad of respiratory insufficiency, acute cardiovascular failure, and coagulopathy.
    [Show full text]
  • The Plasmin–Antiplasmin System: Structural and Functional Aspects
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Bern Open Repository and Information System (BORIS) Cell. Mol. Life Sci. (2011) 68:785–801 DOI 10.1007/s00018-010-0566-5 Cellular and Molecular Life Sciences REVIEW The plasmin–antiplasmin system: structural and functional aspects Johann Schaller • Simon S. Gerber Received: 13 April 2010 / Revised: 3 September 2010 / Accepted: 12 October 2010 / Published online: 7 December 2010 Ó Springer Basel AG 2010 Abstract The plasmin–antiplasmin system plays a key Plasminogen activator inhibitors Á a2-Macroglobulin Á role in blood coagulation and fibrinolysis. Plasmin and Multidomain serine proteases a2-antiplasmin are primarily responsible for a controlled and regulated dissolution of the fibrin polymers into solu- Abbreviations ble fragments. However, besides plasmin(ogen) and A2PI a2-Antiplasmin, a2-Plasmin inhibitor a2-antiplasmin the system contains a series of specific CHO Carbohydrate activators and inhibitors. The main physiological activators EGF-like Epidermal growth factor-like of plasminogen are tissue-type plasminogen activator, FN1 Fibronectin type I which is mainly involved in the dissolution of the fibrin K Kringle polymers by plasmin, and urokinase-type plasminogen LBS Lysine binding site activator, which is primarily responsible for the generation LMW Low molecular weight of plasmin activity in the intercellular space. Both activa- a2M a2-Macroglobulin tors are multidomain serine proteases. Besides the main NTP N-terminal peptide of Pgn physiological inhibitor a2-antiplasmin, the plasmin–anti- PAI-1, -2 Plasminogen activator inhibitor 1, 2 plasmin system is also regulated by the general protease Pgn Plasminogen inhibitor a2-macroglobulin, a member of the protease Plm Plasmin inhibitor I39 family.
    [Show full text]
  • Plasmin (Human) 1.00 Mg
    Plasmin (Human) 1.00 mg Ref#: HPLAS Lot#: xxxxxx Exp. Date: xxxx-xx Store at -10°C to -20°C For Research Use Only Not for Use in Diagnostic Procedures For in vitro use only Description: Plasmin Format: Frozen in 50mM Hepes/ 50 mM sodium acetate/ 50% glycerol/ pH 8.5 Host: Human Storage: Store between -10°C and -20°C Volume: 1 vial containing 0.962 mL Total Protein: 1.00 mg 1% Concentration: 1.04 mg/mL by Absorbance; Extinction Coefficient E 280 = 17.0 Activity: 228.00 nkat/mg Molecular weight: 83000 daltons Plasminogen is synthesized in the liver and circulates in plasma at a concentration of ~200 μg/mL (~2.3 μM). Plasminogen is a single-chain glycoprotein of ~88 kDa that consists of a catalytic domain followed by five kringle structures. Within these kringle structures are four low-affinity lysine binding sites and one high-affinity lysine binding site. It is through these lysine binding sites that plasminogen binds to fibrin and to α2-Antiplasmin. Native Plasminogen (Glu-Plasminogen) exists in two variants that differ in their extent of glycosylation, and each variant has up to six isoelectric forms with respect to sialic acid content, for a total of 12 molecular forms. Activation of Glu-Plasminogen by the Plasminogen activators Urokinase (UPA), or tissue Plasminogen Activator (tPA) occurs by cleavage after residue Arg560 to produce the two-chain active serine protease Plasmin. In a positive feedback reaction, the Plasmin generated cleaves an ~8 kDa peptide from Glu-Plasminogen, producing lys77- Plasminogen which has a higher affinity for Fibrin and when bound is a preferred substrate for Plasminogen activators such as Urokinase.
    [Show full text]
  • 6. Interaction Between the Coagulation and Complement System
    6. Interaction Between the Coagulation and Complement System Umme Amara1, Daniel Rittirsch 1, Michael Flierl 1, Uwe Bruckner 2, Andreas Klos 3, Florian Gebhard1, John D. Lambris4, and Markus Huber-Lang1,* 1Department of Traumatology, Hand-, Plastic-, and Reconstructive Surgery, University Hospital of Ulm, Ulm, Germany, [email protected] 2Division of Experimental Surgery, University Hospital of Ulm, Ulm, Germany, [email protected] 3Department of Medical Microbiology, Medical School Hannover, Hannover, Germany, [email protected] 4Department of Pathology, University of Pennsylvania, 401 Stellar Chance, Philadelphia, PA 19104, USA, [email protected]. Abstract. The complement system as a main column of innate immunity and the coagulation system as a main column in hemostasis undergo massive activation early after injury. Interactions between the two cascades have often been proposed but the precise molecular pathways of this interplay are still in the dark. To elucidate the mechanisms involved, the effects of various coagulation factors on complement activation and generation of anaphylatoxins were investigated and summarized in the light of the latest literature. Own in vitro findings suggest, that the coagulation factors FXa, FXIa and plasmin may cleave both C5 and C3, and robustly generate C5a and C3a (as detected by immunoblotting and ELISA). The produced anaphylatoxins were found to be biologically active as shown by a dose- dependent chemotactic response of neutrophils and HMC-1 cells, respectively. Thrombin did not only cleave C5 (Huber-Lang et al. 2006) but also in vitro-generated C3a when incubated with native C3. The plasmin-induced cleavage activity could be dose-dependently blocked by the serine protease inhibitor aprotinin and leupeptine.
    [Show full text]
  • Activation of the Plasma Kallikrein-Kinin System in Respiratory Distress Syndrome
    003 I-3998/92/3204-043 l$03.00/0 PEDIATRIC RESEARCH Vol. 32. No. 4. 1992 Copyright O 1992 International Pediatric Research Foundation. Inc. Printed in U.S.A. Activation of the Plasma Kallikrein-Kinin System in Respiratory Distress Syndrome OLA D. SAUGSTAD, LAILA BUP, HARALD T. JOHANSEN, OLAV RPISE, AND ANSGAR 0. AASEN Department of Pediatrics and Pediatric Research [O.D.S.].Institute for Surgical Research. University of Oslo [L.B.. A.O.A.], Rikshospitalet, N-0027 Oslo 1, Department of Surgery [O.R.],Oslo City Hospital Ullev~il University Hospital, N-0407 Oslo 4. Department of Pharmacology [H. T.J.],Institute of Pharmacy, University of Oslo. N-0316 Oslo 3, Norway ABSTRAm. Components of the plasma kallikrein-kinin proteins that interact in a complicated way. When activated, the and fibrinolytic systems together with antithrombin 111 contact factors plasma prekallikrein, FXII, and factor XI are were measured the first days postpartum in 13 premature converted to serine proteases that are capable of activating the babies with severe respiratory distress syndrome (RDS). complement, fibrinolytic, coagulation, and kallikrein-kinin sys- Seven of the patients received a single dose of porcine tems (7-9). Inhibitors regulate and control the activation of the surfactant (Curosurf) as rescue treatment. Nine premature cascades. C1-inhibitor is the most important inhibitor of the babies without lung disease or any other complicating contact system (10). It exerts its regulatory role by inhibiting disease served as controls. There were no differences in activated FXII, FXII fragment, and plasma kallikrein (10). In prekallikrein values between surfactant treated and non- addition, az-macroglobulin and a,-protease inhibitor inhibit treated RDS babies during the first 4 d postpartum.
    [Show full text]
  • Assessing Plasmin Generation in Health and Disease
    International Journal of Molecular Sciences Review Assessing Plasmin Generation in Health and Disease Adam Miszta 1,* , Dana Huskens 1, Demy Donkervoort 1, Molly J. M. Roberts 1, Alisa S. Wolberg 2 and Bas de Laat 1 1 Synapse Research Institute, 6217 KD Maastricht, The Netherlands; [email protected] (D.H.); [email protected] (D.D.); [email protected] (M.J.M.R.); [email protected] (B.d.L.) 2 Department of Pathology and Laboratory Medicine and UNC Blood Research Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; [email protected] * Correspondence: [email protected]; Tel.: +31-(0)-433030693 Abstract: Fibrinolysis is an important process in hemostasis responsible for dissolving the clot during wound healing. Plasmin is a central enzyme in this process via its capacity to cleave fibrin. The ki- netics of plasmin generation (PG) and inhibition during fibrinolysis have been poorly understood until the recent development of assays to quantify these metrics. The assessment of plasmin kinetics allows for the identification of fibrinolytic dysfunction and better understanding of the relationships between abnormal fibrin dissolution and disease pathogenesis. Additionally, direct measurement of the inhibition of PG by antifibrinolytic medications, such as tranexamic acid, can be a useful tool to assess the risks and effectiveness of antifibrinolytic therapy in hemorrhagic diseases. This review provides an overview of available PG assays to directly measure the kinetics of plasmin formation and inhibition in human and mouse plasmas and focuses on their applications in defining the role of plasmin in diseases, including angioedema, hemophilia, rare bleeding disorders, COVID- 19, or diet-induced obesity.
    [Show full text]
  • Hemostasis at a Glance ❯❯ Cody Alcott, DVM ❯❯ Charles Brockus, DVM, ❯❯ Brett Sponseller, DVM, Response to Initial David M
    3 CE CREDITS CE Article 2 Hemostasis At a Glance ❯❯ Cody Alcott, DVM ❯❯ Charles Brockus, DVM, ❯❯ Brett Sponseller, DVM, Response to initial David M. Wong, DVM, PhD, DACVIM, DACVP PhD, DACVIM MS, DACVIM Charles River Labs Iowa State University vascular injury Sparks, Nevada Page 78 Iowa State University Platelet plug formation Abstract: Hemostasis, the coagulation cascade, and clot formation present a daunting list of and primary hemostasis Page 79 factors, pathways, and interactions to equine clinicians. A basic knowledge of hemostasis is necessary to evaluate various disease processes in horses. Initial injury to the vascular en- Endothelial cell reactions Page 80 dothelium results in local vasoconstriction and formation of a platelet plug. This initial response reduces blood loss from the damaged vessel but may be inadequate at maintaining hemostasis Secondary hemostasis alone. Therefore, the intrinsic, extrinsic, and common coagulation pathways interact with one Page 80 another to form thrombin and ultimately stabilize the platelet plug. Thrombin accomplishes this The coagulation cascade: by converting soluble fibrinogen to insoluble fibrin, resulting in clot stabilization. Alternatively, the extrinsic pathway fibrinolysis promotes resolution of clot formation. Because of multiple positive and negative Page 80 feedback interactions as well as multiple circulating mediators and inhibitors, coagulation and The coagulation cascade: fibrinolysis are finely controlled systems in healthy patients. However, perturbations of the the intrinsic pathway coagulation cascade in disease states can result in severe complications or death. Page 81 Common pathway lterations in coagulation and Response to Initial Vascular Injury Page 81 fibrinolysis are relatively uncom- The main function of the coagulation sys- Clot formation Amon in horses, but when they tem is to control hemorrhage from the Page 82 occur, especially in critically ill patients, vascular endothelium when it has been T h r o m b i n general knowledge of the associated damaged by trauma or inflammation.
    [Show full text]
  • Reduction of Contact Activation Related Fibrinolytic Activity in Factor XII Deficient Patients
    Reduction of contact activation related fibrinolytic activity in factor XII deficient patients. Further evidence for the role of the contact system in fibrinolysis in vivo. M Levi, … , H R Büller, J W ten Cate J Clin Invest. 1991;88(4):1155-1160. https://doi.org/10.1172/JCI115416. Research Article In this study the contribution of activation of the contact system to activation of the fibrinolytic system in vivo was investigated in healthy volunteers and in factor XII deficient patients. The plasminogen activating activity in plasma from healthy volunteers after infusion of desamino D-arginine vasopressin (DDAVP) was only partially blocked (for 77%) with specific antibodies to tissue-type plasminogen activator and urokinase type plasminogen activator. The residual activity could be quenched by a monoclonal antibody that inhibits factor XII activity and was not present in patients with a factor XII deficiency. The formation of plasmin upon the DDAVP stimulus as reflected by circulating plasmin-alpha 2-antiplasmin complexes was lower in factor XII deficient patients than in healthy volunteers. Activation of the contact system occurred after DDAVP infusion in healthy volunteers and was absent in factor XII deficient patients. These results indicate that DDAVP induces a plasminogen activating activity that is partially dependent on activation of the contact system and that contributes to the overall fibrinolytic activity as indicated by the formation of plasmin-alpha 2-antiplasmin complexes. This fibrinolytic activity is impaired in factor XII deficient patients which may explain the occurrence of thromboembolic complications in these patients. Find the latest version: https://jci.me/115416/pdf Reduction of Contact Activation Related Fibrinolytic Activity in Factor Xll Deficient Patients Further Evidence for the Role of the Contact System in Fibrinolysis In Vivo Marcel Levi,* C.
    [Show full text]
  • Plasmin and Plasminogen System in the Tumor Microenvironment: Implications for Cancer Diagnosis, Prognosis, and Therapy
    cancers Review Plasmin and Plasminogen System in the Tumor Microenvironment: Implications for Cancer Diagnosis, Prognosis, and Therapy Alamelu G. Bharadwaj 1, Ryan W. Holloway 1, Victoria A. Miller 1 and David M. Waisman 1,2,* 1 Department of Pathology, Dalhousie University, Halifax, NS B3H 4R2, Canada; [email protected] (A.G.B.); [email protected] (R.W.H.); [email protected] (V.A.M.) 2 Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS B3H 4R2, Canada * Correspondence: [email protected] Simple Summary: In this review, we present a detailed discussion of how the plasminogen-activation system is utilized by tumor cells in their unrelenting attack on the tissues surrounding them. Plasmin is an enzyme which is responsible for digesting several proteins that hold the tissues surrounding solid tumors together. In this process tumor cells utilize the activity of plasmin to digest tissue barriers in order to leave the tumour site and spread to other parts of the body. We specifically focus on the role of plasminogen receptor—p11 which is an important regulatory protein that facilitates the conversion of plasminogen to plasmin and by this means promotes the attack by the tumour cells on their surrounding tissues. Abstract: The tumor microenvironment (TME) is now being widely accepted as the key contributor Citation: Bharadwaj, A.G.; Holloway, to a range of processes involved in cancer progression from tumor growth to metastasis and chemore- R.W.; Miller, V.A.; Waisman, D.M. sistance. The extracellular matrix (ECM) and the proteases that mediate the remodeling of the ECM Plasmin and Plasminogen System in form an integral part of the TME.
    [Show full text]
  • Activation of the Classical Pathway of Complement by Hageman Factor Fragment*
    CORE Metadata, citation and similar papers at core.ac.uk Provided by PubMed Central ACTIVATION OF THE CLASSICAL PATHWAY OF COMPLEMENT BY HAGEMAN FACTOR FRAGMENT* BY BERHANE GHEBREHIWET, MICHAEL SILVERBERG, AND ALLEN P. KAPLAN From the Department of Medicine, Division of Allergy, Rheumatology, and Clinical Immunology, State University of New York, Stony Brook, New York 11794 There is a striking similarity between the mechanisms involved in the activation of the complement system and those of the Hageman factor-dependent pathways that lead to blood coagulation, fibrinolysis, and kinin formation Both systems require an initial contact with an activator which in turn triggers a cascade of reactions involving the sequential interactions of a series of enzymes and regulators. Furthermore, certain pro_reins seem to operate as regulators of both systems. The serum C1 inactivator (CI-INA), 1 an inhibitor of activated C1 that is absent in patients with hereditary angioedema (1), is known to inhibit the activities ofplasmin (2, 3), activated Ha_geman factor (2-5), anaphylatoxin inactivator (2), Factor XIa (6), kallikrein (2, 7),C lr, and Cls (2, 3) through the formation of stoichiometric complexes. In the absence of C 1-INA, activated Hageman factor may activate the kinin system by conversion of prekallikrein to kallikrein, which in turn cleaves bradykinin from high molecular weight kininogen. Since bradykinin is a potent mediator of enhanced vascular permeability, the pathogenesis of hereditary angioedema may involve interactions between the Hageman factor-dependent pathways and the classical pathway of complement activation. Previous evidence has suggested that C 1 activation in this disease is Hageman factor dependent (8).
    [Show full text]
  • Regulated Proteolytic Processing of Reelin Through Interplay of Tissue Plasminogen Activator (Tpa), ADAMTS-4, ADAMTS-5, and Their Modulators
    Regulated Proteolytic Processing of Reelin through Interplay of Tissue Plasminogen Activator (tPA), ADAMTS-4, ADAMTS-5, and Their Modulators Dimitrije Krstic, Myriam Rodriguez, Irene Knuesel* Institute of Pharmacology and Toxicology, University of Zurich, Zurich, Switzerland Abstract The extracellular signaling protein Reelin, indispensable for proper neuronal migration and cortical layering during development, is also expressed in the adult brain where it modulates synaptic functions. It has been shown that proteolytic processing of Reelin decreases its signaling activity and promotes Reelin aggregation in vitro, and that proteolytic processing is affected in various neurological disorders, including Alzheimer’s disease (AD). However, neither the pathophysiological significance of dysregulated Reelin cleavage, nor the involved proteases and their modulators are known. Here we identified the serine protease tissue plasminogen activator (tPA) and two matrix metalloproteinases, ADAMTS-4 and ADAMTS-5, as Reelin cleaving enzymes. Moreover, we assessed the influence of several endogenous protease inhibitors, including tissue inhibitors of metalloproteinases (TIMPs), a-2-Macroglobulin, and multiple serpins, as well as matrix metalloproteinase 9 (MMP-9) on Reelin cleavage, and described their complex interplay in the regulation of this process. Finally, we could demonstrate that in the murine hippocampus, the expression levels and localization of Reelin proteases largely overlap with that of Reelin. While this pattern remained stable during normal aging, changes in their protein levels coincided with accelerated Reelin aggregation in a mouse model of AD. Citation: Krstic D, Rodriguez M, Knuesel I (2012) Regulated Proteolytic Processing of Reelin through Interplay of Tissue Plasminogen Activator (tPA), ADAMTS-4, ADAMTS-5, and Their Modulators. PLoS ONE 7(10): e47793.
    [Show full text]