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Recombinant production, crystallization and crystal structure determination of dihydroorotate dehydrogenase from Leishmania (Viannia) braziliensis
Author(s) -
Reis Renata Almeida Garcia,
Lorenzato Eder,
Silva Valeria Cristina,
ato Maria Cristina
Publication year - 2015
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x15000886
Subject(s) - dihydroorotate dehydrogenase , pyrimidine metabolism , leishmania braziliensis , biology , biochemistry , enzyme , leishmania , pectobacterium carotovorum , leishmaniasis , genetics , cutaneous leishmaniasis , parasite hosting , gene , purine , world wide web , computer science
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 ( Lb DHODH) was developed and its crystal structure was determined at 2.12 Å 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 Lb DHODH 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 Lb DHODH 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.