Crystal structure of Trypanosoma cruzi dihydroorotate

dehydrogenase from Y strain

Matheus P. Pinheiro a, Jorge Iulek b, M. Cristina Nonato a,*

a Laborato´ rio de Cristalografia de Proteı´nas, Departamento de Fı´sica e Quı´mica, Faculdade de Cieˆncias Farmaceˆuticas de Ribeira˜o Preto,

Universidade de Sa˜o Paulo, Ribeira˜o Preto S.P. 14040-903, Brazil

b Setor de Cieˆncias Exatas e Naturais, Departamento de Quı´mica, Universidade Estadual de Ponta Grossa, Ponta Grossa P.R. 84030-000, Brazil

Received 5 February 2008

Available online 25 February 2008

Abstract

Trypanosoma cruzi is the etiological agent of Chagas’ disease, a pathogenesis that affects millions of  eople in Latin America. Here, we report the crystal structure of dihydroorotate dehydrogenase (DHODH) from T. cruzi strain Y solved at 2.2A ˚ resolution. DHODH is a flavin mononucleotide containing enzyme, which catalyses the oxidation of L-dihydroorotate to orotate, the fourth step and only redox reaction in the de novo biosynthesis of pyrimidine nucleotides. Genetic studies have shown that DHODH is essential for T. cruzi survival, validating the idea that this enzyme can be considered an attractive target for the development of antichagasic drugs. In our work, a detailed analysis of T. cruzi DHODH crystal structure has allowed us to suggest potential sites to be further exploited for the design of highly specific inhibitors through the technology of structure-based drug design.

_ 2008 Elsevier Inc. All rights reserved.

Keywords: Trypanosoma cruzi; Y strain; Dihydroorotate dehydrogenase; Cloning; Expression; Purification; Crystallization; X-ray crystallography