Thromb Haemost 2001; 85(04): 710-715
DOI: 10.1055/s-0037-1615657
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Characterization of Aspercetin, a Platelet Aggregating Component from the Venom of the Snake Bothrops asper which Induces Thrombocytopenia and Potentiates Metalloproteinase-induced Hemorrhage

Alexandra Rucavado
1   Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica
,
Mónica Soto
1   Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica
,
Aura S. Kamiguti
2   Department of Haematology, University of Liverpool, Royal Liverpool University Hospital, Liverpool, UK
,
David G. Theakston
3   Venom Research Unit, Liverpool School of Tropical Medicine, Liverpool, UK
,
Jay W. Fox
4   Department of Microbiology, University of Virginia Health Sciences Center, Charlottesville, Virginia, USA
,
Teresa Escalante
1   Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica
,
José María Gutiérrez
1   Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica
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Publikationsverlauf

Received 10. Juli 2001

Accepted after resubmission 09. November 2000

Publikationsdatum:
08. Dezember 2017 (online)

Summary

Thrombocytopenia occurs in a number of patients bitten by Bothrops asper, a species responsible for the majority of snakebites in Central America and southern Mexico. In this work we describe the isolation of a new platelet-aggregating protein, named aspercetin, from the venom of B. asper, which induces thrombocytopenia in mice. Isolation was carried out by a combination of ion-exchange chromatography on DEAE-Sepharose and affinity chromatography on Affi-Gel Blue. Aspercetin is a disulfide-linked heterodimer, with a pI of 4.5 and a molecular mass of 29,759 Da, detemined by MALDI-ESI mass spectrometry. N-terminal sequence shows homology with a number of venom proteins which belong to the C-type lectin family. Aspercetin has functional similarities with botrocetin, from B. jararaca venom, since it induces platelet aggregation only in the presence of plasma or purified von Willebrand factor. Aspercetin-mediated platelet aggregation results from the interaction of von Willebrand factor with platelet receptor GPIb. Aspercetin lacks anticoagulant effect and does not agglutinate erythrocytes, in contrast with other representatives of the C-type lectin family isolated from snake venoms. Moreover, aspercetin is not lethal, nor does it induce myonecrosis, hemorrhage and edema. When injected intravenously or intramuscularly in mice it induces a rapid, dose-dependent drop in platelet counts and prolongs the bleeding time, suggesting that it may play a role in the thrombocytopenia that develops in a number of B. asper envenomations. Moreover, mice injected intravenously with aspercetin and then receiving an intradermal injection of B. asper hemorrhagic metalloproteinase BaP1 develop a larger hemorrhagic lesion than mice receiving only BaP1. This suggests that aspercetin, by reducing platelet numbers, may contribute to the hemorrhagic effect characteristic of B. asper envenomations.

 
  • References

  • 1 Markland FS. Snake venoms and the hemostatic system. Toxicon 1998; 36: 1749-800.
  • 2 Marsh NA. Snake venoms affecting the haemostatic mechanism – a consideration of their mechanisms, practical applications and biological significance. Blood Coagulation and Fibrinolysis 1994; 5: 399-410.
  • 3 Cardoso JLC, Fan HW, Franca FOS, Jorge MT, Leite RP, Nishioka SA, Avila A, Sano-Martins IS, Tomy SC, Santoro ML, Chudzinski AM, Castro SB, Kamiguti AS, Kelen EMA, Hirata MH, Mirandola RMS, Theakston RDG, Warrell DA. Randomized comparative trial of three antivenoms in the treatment of envenoming by lance-headed vipers (Bothrops jararaca) in São Paulo, Brazil. Q J Med 1993; 86: 315-25.
  • 4 Otero R, Gutiérrez JM, Núñez V, Robles A, Estrada R, Segura E, Toro MF, García ME, Díaz A, Ramírez EC, Gómez G, Castañeda J, Moreno ME. A randomized double-blind clinical trial of two antivenoms in patients bitten by Bothrops atrox in Colombia. Trans Royal Soc Trop Med Hyg 1996; 90: 696-700.
  • 5 Warrell DA. Clinical toxicology of snakebite in Africa and the Middle East/Arabian Peninsule. In: Handbook of Clinical Toxicology of Animal Venoms and Poisons. Meier J, White J. eds. Boca Raton: CRC Press; 1995: 433-92.
  • 6 Niewiarowski S, McLane MA, Kloczewiak M, Stewart GJ. Disintegrins and other naturally occurring antagonists of platelet fibrinogen receptors. Sem Hematol 1994; 31: 289-300.
  • 7 Clemetson KJ, Polgár J, Clemetson JM. Snake venom C-type lectins as tools in platelet research. Platelets 1998; 9: 165-9.
  • 8 Ownby CL, Bjarnason JB, Tu AT. Hemorrhagic toxins from rattlesnake (Crotalus atrox) venom: pathogenesis of hemorrhage induced by three purified toxins. Am J Pathol 1978; 93: 201-18.
  • 9 Bjarnason JB, Fox JW. Hemorrhagic metalloproteinases from snake venoms. Pharmac Ther 1994; 62: 325-72.
  • 10 Gutiérrez JM. Clinical toxicology of snake bites in Central America. In: Handbook of Toxicology of Animal Venoms and Poisons. Meier J, White J. eds. Boca Raton: CRC Press; 1995: 645-65.
  • 11 Lomonte B, Tarkowski A, Hanson LÅ. Host response to Bothrops asper snake venom: analysis of edema formation, inflammatory cells and cytokine release in a mouse model. Inflammation 1993; 17: 93-105.
  • 12 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
  • 13 Cortés-Bratti X, Chaves-Olarte E, Lagergard T, Thelestam M. The cyto-lethal distending toxin from the chancroid bacterium Haemophilus ducreyi induces cell-cycle arrest in the G2 phase. J. Clin. Invest. 1999; 103: 107-15.
  • 14 Kamiguti AS, Hay CR, Zuzel M. Inhibition of collagen-induced platelet aggregation as the result of cleavage of alpha 2 beta 1-integrin by the snake venom metalloproteinase jararhagin. Biochem. J. 1996; 320: 635-41.
  • 15 Lomonte B, Rojas G, Gutiérrez JM, Ramírez G. Isolation of a galactose-binding lectin from the venom of the snake Bothrops godmani (Godmann’s pit viper). Toxicon 1990; 28: 75-81.
  • 16 Atoda H, Morita T. A novel blood coagulation factor IX/factor X-binding protein with anticoagulant activity from the venom of Trimeresurus flavoviridis (habu snake): isolation and characterization. J. Biochem. 1989; 106: 808-13.
  • 17 Gutiérrez JM, Lomonte B, Chaves F, Moreno E, Cerdas L. Pharmacological activities of a toxic phospholipase A isolated from the venom of the snake Bothrops asper . Comp Biochem Physiol 1986; 84C: 159-64.
  • 18 Gutiérrez JM, Gené JA, Rojas G, Cerdas L. Neutralization of proteolytic and hemorrhagic activities of Costa Rican snake venoms by a polyvalent antivenom. Toxicon 1985; 23: 887-93.
  • 19 Gutiérrez JM, Arroyo O, Bolaños R. Mionecrosis, hemorragia y edema inducidos por el veneno de Bothrops asper en ratón blanco. Toxicon 1980; 18: 603-10.
  • 20 Denis C, Methia N, Frenette PS, Rayburn H, Ullman-Cullere M, Hynes RO, Wagner DD. A mouse model of severe von Willebrand disease: defects in hemostasis and thrombosis. Proc. Natl. Acad. Sci USA 1998; 95: 9524-9.
  • 21 Gutiérrez JM, Romero M, Díaz C, Borkow G, Ovadia M. Isolation and characterization of a metalloproteinase with weak hemorhagic activity from the venom of the snake Bothrops asper (terciopelo). Toxicon 1995; 33: 19-29.
  • 22 Rucavado A, Núñez J, Gutiérrez JM. Blister formation and skin damage induced by BaP1, a haemorrhagic metalloproteinase from the venom of the snake Bothrops asper . Int. J. Exp. Path. 1998; 79: 245-54.
  • 23 Andrews RK, Booth WJ, Gorman JJ, Castaldi PA, Berndt MC. Purification of botrocetin from Bothrops jararaca venom. Analysis of the botrocetin-mediated interaction between von Willebrand factor and the human platelet membrane glycoprotein Ib-IX complex. Biochemistry 1989; 28: 8317-26.
  • 24 Ozeki Y, Matsui T, Hamako J, Suzuki M, Fujimura Y, Yoshida E, Nishida S, Titani K. C-type galactoside-binding lectin from Bothrops jararaca venom: comparison of its structure and function with those of botrocetin. Arch. Biochem. Biophys. 1994; 308: 306-10.
  • 25 Sekiya F, Atoda H, Morita T. Isolation and characterization of an anticoagulant protein homologous to botrocetin from the venom of Bothrops jararaca . Biochemistry 1993; 32: 6892-7.
  • 26 Atoda H, Morita T. A novel blood coagulation factor IX/factor X-binding protein with anticoagulant activity from the venom of Trimeresurus flavoviridis (habu snake): isolation and characterization. J Biochem 1989; 106: 808-13.
  • 27 Kawasaki T, Fujmura Y, Usami Y, Suzuki M, Miura S, Sakurai Y, Makita K, Taniuchi Y, Hirano K, Titani K. Complete amino acid sequence and identification of the platelet glycoprotein Ib-binding site of jararaca GPIb-BP, a snake venom protein isolated from B. jararaca . J Biol Chem 1996; 271: 10635-9.
  • 28 Kowalska MA, Tau L, Holt JC, Peng M, Karczewski J, Calvete JJ, Niewarowski S. Alboaggregins A. and B. Structure and interaction with human platelets. Tromb Haemost 1998; 79: 609-13.
  • 29 Hamako J, Matsui T, Suzuki M, Ito M, Makita K, Fujimura Y, Ozeki Y, Titani K. Purification and characterization of bitiscetin, a novel von Willebrand factor modulator protein from Bitis arietans snake venom. Biochem Biophys Res Comm 1996; 226: 273-9.
  • 30 Brinkhous KM, Barnes DS, Potter JY, Read MS. Von Willebrand syndrome induced by a Bothrops venom factor: bioassay for venom coagglutinin. Proc Natl Acad Sci USA 1981; 78: 3230-4.
  • 31 Sanders WE, Read MS, Reddick RL, Garris JB, Brinkhous KM. Thrombotic thrombocytopenia with von Willebrand factor deficiency induced by botrocetin. An animal model. Lab Invest 1988; 59: 443-52.
  • 32 Rucavado A, Lomonte B, Ovadia M, Gutiérrez JM. Local tissue damage induced by BaP1, a metalloproteinase isolated from Bothrops asper (terciopelo) snake venom. Exp Molec Pathol 1995; 63: 186-99.
  • 33 Moreira L, Borkow G, Ovadia M, Gutiérrez JM. Pathological changes induced by BaH1, a hemorrhagic metalloproteinase isolated from Bothrops asper (terciopelo) snake venom, on mouse capillary blood vessels. Toxicon 1994; 32: 977-87.