Caracterização bioquímica e funcional de uma nova metaloproteinase isolada da peçonha de Bothropoides pauloensis (bothrops pauloensis)

• Ano de defesa: 2011
• Autor(a) principal: Souza, Dayane Lorena Naves de
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Uberlândia; BR; Programa de Pós-graduação em Genética e Bioquímica; Ciências Biológicas; UFU
• Palavras-chave em Português: Bothropoides pauloensis; Metaloprotease; Não hemorrágica; Peçonha de serpentes; Anticorpos; Serpente peçonhenta - Peçonha; Bothrops; Metalloproteinase; Snake venom; Antibodies; CNPQ::CIENCIAS BIOLOGICAS::GENETICA
• Link de acesso: https://repositorio.ufu.br/handle/123456789/15871

Chapter II: In the present study, a metalloproteinase named BpMPI was isolated from Bothropoides pauloensis snake venom and its biochemical, enzymatic and immunochemical characteristics were determined. BpMPI was purified in two chromatography steps on ion exchange resins CM-Sepharose Fast flow and Sephacryl S-300. This protease was homogeneous on SDS-PAGE and showed a single chain polypeptide of 23 kDa under reducing conditions. The primary structure of the enzyme showed high similarity with other SVMPs enzymes from snake venoms. BpMPI showed proteolytic activity upon azocasein and bovine fibrinogen and was inhibited by EDTA, 1,10 phenantroline and β-mercaptoethanol. Moreover, this enzyme showed stability at neutral and alkaline pH and it was inactivated at high temperatures. BpMPI was able to hidrolyse glandular and tecidual kallikrein substrates, but was unable to act upon factor Xa and plasmin substrates. The enzyme did not able to induce local hemorrhage in the dorsal region of mice even at high doses. Immunochemistry studies showed that BpMPI was high immunogenic and the antibodies anti-BpMP-I were very efficient to neutralize the hemorrhagic activity and systemic alterations induced by Bothropoides pauloensis snake venom.