Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of recombinant human fumarase

Autor(es)

Pereira de Padua, Ricardo Augusto ; Nonato, Maria Cristina

• Tipo de documento: Artigo Científico
• Fonte: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS; v. 70, p. 3-pg., 2014-01-01.

Resumo

Human fumarase (HsFH) is a well-known citric acid cycle enzyme and is therefore a key component in energy metabolism. Genetic studies on human patients have shown that polymorphisms in the fumarase gene are responsible for diseases such as hereditary leiomyomatosis and renal cell cancer. As a first step in unravelling the molecular basis of the mechanism of fumarase deficiency in genetic disorders, the HsFH gene was cloned in pET-28a, heterologously expressed in Escherichia coli, purified by nickel-affinity chromatography and crystallized using the vapour-diffusion technique. X-ray diffraction experiments were performed at a synchrotron source and the structure was solved at 2.1 angstrom resolution by molecular replacement. (AU)

• Processo FAPESP: 08/08262-6 - Caracterização cinética, estrutural e funcional dos genes LmjF24.0320. e LmjF29.1960 que codificam a enzima fumarato hidratase em Leishmania major
• Beneficiário: Maria Cristina Nonato
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 08/11644-8 - Caracterização estrutural e funcional das isoformas da enzima fumarato hidratase de Trypanosoma cruzi
• Beneficiário: Ricardo Augusto Pereira de Pádua
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto