Archives of Toxicology

, Volume 87, Issue 1, pp 197–208 | Cite as

Isolation and characterization of two new non-hemorrhagic metalloproteinases with fibrinogenolytic activity from the mapanare (Bothrops colombiensis) venom

  • María E. Girón
  • Alexis Rodríguez-Acosta
  • Ana María Salazar
  • Elda E. Sánchez
  • Jacob Galán
  • Carlos Ibarra
  • Belsy Guerrero
Biologicals

Abstract

Colombienases are acidic, low molecular weight metalloproteinases (Mr of 23,074.31 Da colombienase-1 and 23,078.80 Da colombienase-2; pI of 6.0 and 6.2, respectively) isolated from Bothrops colombiensis snake venom. The chromatographic profile in RP-HPLC and its partial sequence confirmed its high homogeneity. Both colombienases present fibrino(geno)lytic activity, but did not show any hemorrhagic, amidolytic, plasminogen activator or coagulant activities, and no effect on platelet aggregation induced by collagen or ADP. Both enzymes were strongly active on fibrinogen Aα chains followed by the Bβ chains, and colombienases-2, at high doses, also degraded the γ chains. This activity was stable at temperatures ranging between 4 and 37 °C, with a maximum activity at 25 °C, and at pHs between 7 and 9. The homology demonstrated by the comparison of sequences, with zinc-dependent metalloproteinases, as well as the metal chelant effects on, confirmed that the colombienases were metalloproteinases, particularly to α-fibrinogenases belonging to the P-I class of SVPMs (20–30 kDa), which contain only the single-domain proteins. The biological characteristics of the colombienases confer a therapeutic potential, since they contain a high fibrino(geno)lytic activity, devoid of hemorrhagic activity. These metalloproteinases might be explored as thrombolytic agents given that they dissolve fibrin clots or prevent their formation.

Keywords

Bothrops colombiensis Colombienases Fibrinolytic activity Fibrinogenolytic activity Metalloproteinases Hemostasis 

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • María E. Girón
    • 1
  • Alexis Rodríguez-Acosta
    • 1
  • Ana María Salazar
    • 2
  • Elda E. Sánchez
    • 3
  • Jacob Galán
    • 4
  • Carlos Ibarra
    • 2
  • Belsy Guerrero
    • 2
    • 5
  1. 1.Laboratorio de Inmunoquímica y UltraestructuraInstituto Anatómico de la Universidad Central de VenezuelaCaracasVenezuela
  2. 2.Laboratorio de Fisiopatología, Centro de Medicina ExperimentalInstituto Venezolano de Investigaciones Científicas (IVIC)CaracasVenezuela
  3. 3.Department of Chemistry and the National Natural Toxins Research CenterTexas A&M University-KingsvilleKingsvilleUSA
  4. 4.Department of Molecular Biology, Institute for Research in Immunology and Cancer (IRIC)Université de MontréalMontrealCanada
  5. 5.Laboratorio de Fisiopatología y Coagulación SanguíneaInstituto Venezolano de Investigaciones CientíficasCaracasVenezuela

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