Article
Isolation and characterization of two new non-hemorrhagic metalloproteinases with fibrinogenolytic activity from the mapanare (Bothrops colombiensis) venom
Autor
Giron, Marıa E
Rodríguez-Acosta, Alexis
Salazar, Ana Marıa
Sanchez, Elda E
Galan, Jacob
Ibarra, Carlos
Guerrero, Belsy
Institución
Resumen
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 Aa chains followed
by the Bb chains, and colombienases-2, at high doses,
also degraded the c 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 a-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. Financial support was provided by the Science and Technology Foundation (FONACIT,
Grant PG-2005000400), Venezuela; NCRR Viper Resource grant (P40 RR018300-09), Kingsville, TX, USA; and the Instituto Venezolano de Investigaciones Cientı´ficas, Caracas, Venezuela.