Descripción de Hutmannin-1, una nueva metaloproteasa piii del veneno de la serpiente neotropical mapanare de lansberg (Porthidium lansbergii Hutmanni) con actividades fibrino (geno) líticas y hemorrágicas

Description of Hutmannin-1, a new piii-metalloprotease from the venom of the neotropical lansberg’s hognose Viper (Porthidium lansbergii hutmanni) With fibrino(geno) lytic and haemorrhagic activities

• María Eugenia Pineda Universidad Central de Venezuela
• Jonás Perales Laboratory of Toxinology, Oswaldo Cruz Institute
• Elda Eliza Sánchez Texas A&M University-Kingsville
• Tomas Hermoso Universidad Central de Venezuela
• María Narvaez Universidad Central de Venezuela
• Alexander Chapeaurouge Laboratory of Toxinology, Oswaldo Cruz Institute
• Alba Marlene Vargas Universidad Central de Venezuela
• Alexis Rodríguez-Acosta Universidad Central de Venezuela

Palabras clave: Anticoagulante, antitoxina, coagulación, hemostasia, hemorragia, veneno de serpiente

Resumen

El objetivo de este trabajo fue caracterizar hutmannin-1 (hut-1), una nueva metaloproteasa de clase P-III de ~ 62 kDa de la serpiente Porthidium lansbergii hutmanni (P.l.h ) (Isla Margarita, Venezuela). Para caracterizar esta proteína, el veneno crudo de P.l.h se fraccionó mediante cromatografía de exclusión molecular, cromatografía de intercambio aniónico y cromatografía líquida del alto rendimiento (HPLC). Hutmannin-1 se identificó por espectrometría de masas MALDI-TOF / TOF, y el veneno se analizó por SDS-PAGE. Se determinó la letalidad, la dosis hemorrágica mínima (MHD), el efecto de la temperatura sobre la actividad, la actividad procoagulante en el plasma humano y las actividades anticoagulantes, desfibrinantes, fibrinolíticos gelatinolíticos y fibrinogenolíticos y de agregación plaquetaria de hut-1. Se realizaron ensayos de reconocimiento antigénico en P.l.h veneno crudo y hut-1 mediante suero anti-ofídico polivalente (PAOS) venezolano. Hut-1 tuvo una fuerte actividad fibrinolítica y fibrinolítica moderada. Estas actividades y la actividad hemorrágica de hut-1 fueron completamente inhibidas por ethylenediaminetetraacetic acid (EDTA). El veneno crudo de P.l.h tuvo una potente actividad anticoagulante en plasma recalcificado e inhibió la agregación plaquetaria inducida por trombina, ADP, colágeno y ristocetina. En contraste, la inhibición de la agregación plaquetaria, actividad anticoagulante y coagulante de hut-1 no se observó con ninguno de los agonistas. Este resultado sugiere que otras proteínas en el veneno crudo, afectan notablemente las funciones plaquetarias y / o los factores de coagulación. El antiveneno comercial venezolano mostró una capacidad limitada para neutralizar la actividad hemorrágica de la hut-1.

Citas

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