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

Autor(es)

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

• Tipo de documento: Artigo Científico
• Fonte: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS; v. 71, p. 6-pg., 2015-05-01.

Resumo

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)

• Processo FAPESP: 11/23504-9 - Exploração da relação estrutura-função da Enzima DHODH no desenvolvimento de novas moléculas com ação tripanocida e leishmanicida
• Beneficiário: Renata Almeida Garcia Reis
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 14/01257-8 - Desenvolvimento de uma plataforma modelo para a busca de ligantes com potencial leishmanicida baseado na inibição seletiva da enzima diidroorotato desidrogenase
• Beneficiário: Eder Lorenzato Junior
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 12/25075-0 - Desenvolvimento de moléculas com ação leishmanicida baseado na inibição seletiva da enzima diidroorotato desidrogenase
• Beneficiário: Maria Cristina Nonato
• Modalidade de apoio: Auxílio à Pesquisa - Regular