Expressão e caracterização bioquímica parcial de uma serinoprotease recombinante da peçonha de Bothrops pauloensis

• Ano de defesa: 2014
• Autor(a) principal: Costa, Guilherme Nunes Moreira
• 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: Bothrops pauloensis; Peçonha de serpente; Thrombin-like; Serinoprotease; Expressão heteróloga; Serpente peçonhenta - Peçonha; Bothrops; Snake venom; Serine peptidase; Heterologous expression; CNPQ::CIENCIAS BIOLOGICAS::GENETICA
• Link de acesso: https://repositorio.ufu.br/handle/123456789/15888; https://doi.org/10.14393/ufu.di.2014.366

CHAPTER II: The snake venom is composed by a diversity of biomolecules with many actions on physiological processes. The serine peptidases are a group of proteases present in the constitution of the venom, capable of interfering on several points of hemostasis. Some serine peptidases has thrombin-like activity, what makes them targets on the development of therapeutics agentes on the treatment of many hemostatic disorders. In this study, a recombinant thrombin-like serine peptidase called rBpSP-II was obtained from the cDNA of the venom gland of the snake Bothrops pauloensis and biochemically characterized. The cDNA correspondent to rBpSP-II was cloned on the pPICZαA vector and inserted on the methylotrophic yeast Pichia pastoris KM71H for the heterologous expression. This enzyme showed single band when analised on SDS-PAGE with approximated molecular mass of 44,5 kDa under reducing conditions. The enzyme rBpSP-II showed clotting activity on bovine plasma and proteolytic activity on fibrinogen, cleaving exclusively the Aα chain. The evaluation of rBpSP-II activity on chromogenic substrates showed that the enzyme has thrombin-like activity due to its capacity to hydrolyze the thrombin substrate.