Recombinant production, crystallization and crystal structure determination of dihydroorotate dehydrogenase from Leishmania (Viannia) braziliensis

Author(s)

Garcia Reis, Renata Almeida ; Lorenzato, Eder, Jr. ; Silva, Valeria Cristina ; Nonato, Maria Cristina

• Document type: Journal article
• Source: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS; v. 71, p. 6-pg., 2015-05-01.

Abstract

The enzyme dihydroorotate dehydrogenase (DHODH) is a flavoenzyme that catalyses the oxidation of dihydroorotate to orotate in the de novo pyrimidine-biosynthesis pathway. In this study, a reproducible protocol for the heterologous expression of active dihydroorotate dehydrogenase from Leishmania (Viannia) braziliensis (LbDHODH) was developed and its crystal structure was determined at 2.12 angstrom resolution. L. (V.) braziliensis is the species responsible for the mucosal form of leishmaniasis, a neglected disease for which no cure or effective therapy is available. Analyses of sequence, structural and kinetic features classify LbDHODH as a member of the class 1A DHODHs and reveal a very high degree of structural conservation with the previously reported structures of orthologous trypanosomatid enzymes. The relevance of nucleotide-biosynthetic pathways for cell metabolism together with structural and functional differences from the respective host enzyme suggests that inhibition of LbDHODH could be exploited for antileishmanicidal drug development. The present work provides the framework for further integrated in vitro, in silico and in vivo studies as a new tool to evaluate DHODH as a drug target against trypanosomatid-related diseases. (AU)

• FAPESP's process: 11/23504-9 - Exploring structure-function relationship of DHODH enzyme in the development of new molecules with trypanocidal and leishmanicidal activities
• Grantee: Renata Almeida Garcia Reis
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 14/01257-8 - Development of a model plataform for the search of leishmanicidal compounds based on the selective inhibition of dihydroorotate dehydrogenase
• Grantee: Eder Lorenzato Junior
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 12/25075-0 - Development of leishmanicidal drugs based on the selective inhibition of the enzyme dihydroorotate dehydrogenase
• Grantee: Maria Cristina Nonato
• Support Opportunities: Regular Research Grants