Laboratório de Cristalografia de Proteínas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
Laboratório de Cristalografia de Proteínas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
Biochimie. 2019 Mar;158:180-190. doi: 10.1016/j.biochi.2019.01.006. Epub 2019 Jan 19.
Trematode worms of the genus Schistosoma are the causing agents of schistosomiasis, a parasitic disease responsible for a considerable economic and healthy burden worldwide. In the present work, the characterization of the enzyme dihydroorotate dehydrogenase from Schistosoma mansoni (SmDHODH) is presented. Our studies demonstrated that SmDHODH is a member of class 2 DHODHs and catalyzes the oxidation of dihydroorotate into orotate using quinone as an electron acceptor by employing a ping-pong mechanism of catalysis. SmDHODH homology model showed the presence of all structural features reported for class 2 DHODH enzymes and reveal the presence of an additional protuberant domain predicted to fold as a flexible loop and absent in the other known class 2 DHODHs. Molecular dynamics simulations showed that the ligand-free forms of SmDHODH and HsDHODH undergo different rearrangements in solution. Well-known class 2 DHODH inhibitors were tested against SmDHODH and HsDHODH and the results suggest that the variable nature of the quinone-binding tunnel between human and parasite enzymes, as well as the differences in structural plasticity involving rearrangements of the N-terminal α-helical domain can be exploited for the design of SmDHODH selective inhibitors, as a strategy to validate DHODH as a drug target against schistosomiasis.
曼氏血吸虫属的吸虫类蠕虫是血吸虫病的病原体,这种寄生虫病在全球范围内造成了相当大的经济和健康负担。在本工作中,描述了曼氏血吸虫(Schistosoma mansoni)二氢乳清酸脱氢酶(SmDHODH)的酶特性。我们的研究表明,SmDHODH 是 2 类 DHODH 酶的成员,通过使用醌作为电子受体,采用乒乓机制的催化,将二氢乳清酸氧化成乳清酸盐。SmDHODH 同源模型显示存在所有报告的 2 类 DHODH 酶的结构特征,并揭示存在一个额外的突出结构域,预测为折叠的柔性环,而在其他已知的 2 类 DHODHs 中不存在。分子动力学模拟表明,配体非结合形式的 SmDHODH 和 HsDHODH 在溶液中经历不同的重排。对 SmDHODH 和 HsDHODH 进行了已知的 2 类 DHODH 抑制剂的测试,结果表明,人源和寄生虫酶之间的醌结合隧道的可变性质,以及涉及 N 端α-螺旋结构域重排的结构可塑性差异,可以被利用来设计 SmDHODH 选择性抑制剂,作为验证 DHODH 作为抗血吸虫病药物靶点的策略。