Crystallization and X Ray Diffraction Data Analyses of the Enzyme Urocanate Hydratase from Trypanosoma cruzi

Author(s)

Boreiko, S. ^[1] ; Silva, M. ^[2] ; Iulek, J. ^[1]

Affiliation

^[1] Univ Estadual Ponta Grossa, Dept Quim, Lab Purificacao Prot, Campus Uvaranas, Ave Gen Carlos Cavalcanti 4748, BR-84030900 Ponta Grossa, PR - Brazil

^[2] Univ Tecnol Fed Parana, Dept Ensino, Campus Ponta Grossa Ave Monteiro Lobato S-N Km 04, BR-84016210 Ponta Grossa, PR - Brazil

• Document type: Journal article
• Source: REVISTA VIRTUAL DE QUIMICA; v. 8, n. 3, p. 678-686, 2016.
• Web of Science Citations: 0

Abstract

Trypanosoma cruzi is the only trypanosomatid pathogenic to humans in which the four enzymes of the histidine degradation pathway were found up to now. The enzyme urocanate hydratase from Trypanosoma cruzi catalyzes the second step of histidine degradation and is essential for the catabolic conversion of urocanate to 4-imidazolone-5-propionate, which finally yields glutamate and further glutamine and tricarboxylic acid cyclr intermediates such as a-ketoglutarate. The recombinant Cterminally His6-tagged protein was expressed in Escherichia coli, purified in a homogenous form and crystallized in several conditions, with the best crystals obtained with 0.04 M dipotassium hydrogen phosphate, 16.00 % (w/v) polyethylene glycol 8,000 and 22.00 % (v/v) glycerol. X ray diffraction data were collected to 2.61 angstrom resolution. The crystals belong to the monoclinic space group P21, with unit cell parameters a = 85.12, b = 137.63, c = 117.69 angstrom, B = 94.69 angstrom and are suitable for molecular replacement phasing. The asymmetric unit contains four monomers, with a V-M of 2.3 angstrom(3)/Da and a solvent content of 45.9 %. (AU)

• FAPESP's process: 08/57910-0 - National Institute of Structural Biotechnology and Medicinal Chemistry in Infectious Diseases
• Grantee: Richard Charles Garratt
• Support Opportunities: Research Projects - Thematic Grants