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

Full text
Author(s):	Pereira de Padua, Ricardo Augusto ; Nonato, Maria Cristina Total Authors: 2
Document type:	Journal article
Source:	ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS; v. 70, p. 3-pg., 2014-01-01.
Abstract
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)

FAPESP's process:	08/08262-6 - Kinetic, Structural and functional characterization of LmjF24.0320. e LmjF29.1960 genes that code for fumarate hydratase in Leishmania major
Grantee:	Maria Cristina Nonato
Support Opportunities:	Regular Research Grants


FAPESP's process:	08/11644-8 - Structural and functional characterization of Trypanosoma cruzi fumarate hydratase isoforms
Grantee:	Ricardo Augusto Pereira de Pádua
Support Opportunities:	Scholarships in Brazil - Doctorate (Direct)