Reis Renata Almeida Garcia, Lorenzato Eder, Silva Valeria Cristina, Nonato Maria Cristina
Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Avenida Café S/N, 14040-903 Ribeirão Preto-SP, Brazil.
Acta Crystallogr F Struct Biol Commun. 2015 May;71(Pt 5):547-52. doi: 10.1107/S2053230X15000886. Epub 2015 Apr 21.
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 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 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.
二氢乳清酸脱氢酶（DHODH）是一种黄素酶，在从头嘧啶生物合成途径中催化二氢乳清酸氧化为乳清酸。在本研究中，开发了一种可重复的方案用于巴西利什曼原虫（Viannia亚属）（LbDHODH）活性二氢乳清酸脱氢酶的异源表达，并以2.12 Å的分辨率确定了其晶体结构。巴西利什曼原虫（Viannia亚属）是导致黏膜利什曼病的病原体，这是一种被忽视的疾病，目前没有治愈方法或有效治疗手段。对序列、结构和动力学特征的分析将LbDHODH归类为1A类DHODHs的成员，并揭示其与先前报道的直系同源锥虫酶结构具有高度的结构保守性。核苷酸生物合成途径对细胞代谢的相关性以及与各自宿主酶的结构和功能差异表明，抑制LbDHODH可用于开发抗利什曼原虫药物。目前的工作为进一步整合体外、计算机模拟和体内研究提供了框架，作为评估DHODH作为抗锥虫相关疾病药物靶点的新工具。
