BmooMPα-III: uma metaloprotease purificada da peçonha da serpente Bothrops moojeni

• Ano de defesa: 2014
• Autor(a) principal: Silva, Thalita Kristhina Alves
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Uberlândia; BR; Programa de Pós-graduação em Biologia Celular e Estrutural Aplicadas; Ciências Biomédicas; UFU
• Palavras-chave em Português: Bothrops moojeni; Metaloprotease; BmooMPα-III; Citologia; Cobra - Veneno; Serpente peçonhenta; CNPQ::CIENCIAS BIOLOGICAS::MORFOLOGIA::CITOLOGIA E BIOLOGIA CELULAR
• Link de acesso: https://repositorio.ufu.br/handle/123456789/12402; https://doi.org/10.14393/ufu.di.2014.191

Proteolytic enzymes from bothrops venom are responsible for the majority of local and systemic effects observed during the envenomation. In this work we describe the purification and characterization of a novel metalloproteinase from Bothrops moojeni snake venom. The enzyme, denoted BmooMPα-III, was purified by a combination of ion exchange (DEAE Sephacel), molecular exclusion (Shepadex G-75) and affinity (HiTrap Chelating HP) chromatographies. BmooMPα-III enzyme represents about 0.75% of the crude venom and shows a single band with an apparent molecular mass of 26 kDa when analyzed by SDS-PAGE, under reducing conditions. BmooMPα-III cleaves first the Aα chain, then βγ, but does not hydrolyzes the γ chain of bovine fibrinogen. Inhibition of fibrinogenolytic activity by metal ion chelating agent such as EDTA allow classifies BmooMPα-III as a metalloprotease. The enzyme was inhibited by β-mercaptoethanol, however, inhibitors such as, aprotinin, leupeptin and PMSF did not Affect the fibrinogenolytic activity. The enzyme is inactive at high temperatures (≥ 60 ° C) and shows maximum activity at pH around 7.0 to 10.0. BmooMPα-III cause toxic effect on liver and kidney cells, but does not alter the cells of the lungs and heart. Preliminary results show that the enzyme BmooMPα-III induces a drastic reduction in HeLa cells and it is able to cause desfibrinating in mice when administered ip. Therefore, it is expected that this enzyme may be of medical importance as a therapeutic agent in the treatment of thrombotic disorders and cancer.