Purification, crystallization and preliminary X-ray diffraction analysis of the glyoxalase II from Leishmania infantum

• Autor(a) principal: Barata, Lauro Euclides Soares
• Data de Publicação: 2006
• Outros Autores: Silva, Marta Sousa, Trincão, José Pedro da Silva, Carvalho, Sandra, Ferreira, António Eduardo N., Bonifácio, Cecília S., Cordeiro, Carlos, Tomás, Ana M., Freire, Ana Ponces, Romão, Maria João
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10362/62936

In trypanosomatids, trypanothione replaces glutathione in all glutathione-dependent processes. Of the two enzymes involved in the glyoxalase pathway, glyoxalase I and glyoxalase II, the latter shows absolute specificity towards trypanothione thioester, making this enzyme an excellent model to understand the molecular basis of trypanothione binding. Cloned glyoxalase II from Leishmania infantum was overexpressed in Escherichia coli, purified and crystallized. Crystals belong to space group C222(1) (unit-cell parameters a = 65.6, b = 88.3, c = 85.2 angstrom) and diffract beyond 2.15 angstrom using synchrotron radiation. The structure was solved by molecular replacement using the human glyoxalase II structure as a search model. These results, together with future detailed kinetic characterization using lactoyltrypanothione, should shed light on the evolutionary selection of trypanothione instead of glutathione by trypanosomatids.