Production, characterization and structural analysis of proteins from Corynebacterium pseudotuberculosis and snake venoms

• Ano de defesa: 2015
• Autor(a) principal: Masood, Rehana [UNESP]
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual Paulista (Unesp)
• Palavras-chave em Português: Biologia molecular; Biofísica; Corynebacterium pseudotuberculosis; Proteínas - Estrutura; Serpente peçonhenta - Peçonha; Cristalografia; Molecular biology
• Link de acesso: http://hdl.handle.net/11449/138506

Corynebacterium pseudotuberculosis (C. pseudotuberculosis) is the etiological agent of Caseous Lymphadenitis (CLA), a disease that affects different groups of animals worldwide, specifically sheep and goat production areas, which result a significant economic losses. These bacteria even infect humans; to date 25 different cases of infections in humans are reported in the literature. Currently, no efficient treatment for this disease is available neither a crystallographic structure of any protein from C. pseudotuberculosis that could assist in better understanding of the mechanism of action of this pathogen. In the present study, we have expressed and purified key proteins that play a major role in the metabolism of this pathogen. Plasmids of three different proteins from C. pseudotuberculosis were designed. Triose phosphate isomerase (TIM) is an active enzyme of glycolytic pathway, which is one of the main energy supplier enzymes for the organisms. This enzyme was expressed, purified and crystallized, the crystal structure was determined at 2.5 Å. Thioredoxin reductase (TrxR), and Thioredoxin (Trx) are the part of thioredoxin (redox) system, in which the thioredoxin act as a substrate for the TrxR. These enzymes have a diverse function in organisms. These enzymes were expressed, purified and characterized. Snake venom proteins were also investigated in this study; five different proteins from genus Bothrops were purified and crystallized. The crystal structure of Atroxlysin-I was determined at 1.8 Å resolutions and compared with different metalloproteinases-I, deposited to PDB