Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom

Autor(es)

Torres-Huaco, Frank Denis ; Werneck, Claudio C. ; Vicente, Cristina Pontes ; Vassequi-Silva, Talita ; Coelho Nery-Diez, Ana Claudia ; Mendes, Camila B. ; Antunes, Edson ; Marangoni, Sergio ; Damico, Daniela C. S.

• Tipo de documento: Artigo Científico
• Fonte: BIOMED RESEARCH INTERNATIONAL; v. 2013, p. 12-pg., 2013-01-01.

Resumo

We report a rapid purification method using one-step chromatography of SVSP Rhombeobin (LMR-47) from Lachesis muta rhombeata venom and its procoagulant activities and effects on platelet aggregation. The venom was fractionated by a single chromatographic step in RP-HPLC on a C8 Discovery BIO Wide Pore, showing high degree of molecular homogeneity with molecular mass of 47035.49 Da. Rhombeobin showed amidolytic activity upon BA rho NA, with a broad optimum pH (7-10) and was stable in solution up to 60 degrees C. The amidolytic activity was inhibited by serine proteinase inhibitors and reducing agents, but not chelating agents. Rhombeobin showed high coagulant activity on mice plasma and bovine fibrinogen. The deduced amino acid sequence of Rhombeobin showed homology with other SVSPs, especially with LM-TL (L. m. muta) and Gyroxin (C. d. terrificus). Rhombeobin acts, in vitro, as a strong procoagulant enzyme on mice citrated plasma, shortening the APTT and PT tests in adose-dependent manner. The protein showed, "ex vivo", a strong defibrinogenating effect with 1 mu g/animal. Lower doses activated the intrinsic and extrinsic coagulation pathways and impaired the platelet aggregation induced by ADP. Thus, this is the first report of a venom component that produces a venom-induced consumptive coagulopathy (VICC). (AU)

• Processo FAPESP: 09/02299-8 - Estudos de estrutura-função de toxinas do veneno de Lachesis muta rhombeata ativas na formação de trombos in vivo
• Beneficiário: Daniela Carla da Silva Damico
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado