Physics Institute of São Carlos, University of São Paulo, São Paulo, SP, Brazil.
Laboratory Medicine & Pathobiology, University of Toronto. 1 King's College Circle, Toronto, Ontario, Canada.
Methods Mol Biol. 2020;2151:159-172. doi: 10.1007/978-1-0716-0635-3_13.
Dihydrofolate reductase (DHFR) is an essential enzyme for nucleotide metabolism used to obtain energy and structural nucleic acids. Schistosoma mansoni has all the pathways for pyrimidine biosynthesis, which include the thymidylate cycle and, consequentially, the DHFR enzyme. Here, we describe the characterization of Schistosoma mansoni DHFR (SmDHFR) using isothermal titration calorimetry for the enzymatic activity and thermodynamic determination, also the folate analogs inhibition. Moreover, X-ray crystallography was used to determine the enzyme atomic model at 1.95 Å.
二氢叶酸还原酶（DHFR）是核苷酸代谢中获取能量和结构核酸所必需的酶。曼氏血吸虫具有嘧啶生物合成的所有途径，包括胸苷酸循环，因此也包括 DHFR 酶。在这里，我们使用等温滴定量热法对酶活性和热力学进行了测定，同时还对叶酸类似物的抑制作用进行了研究，从而描述了曼氏血吸虫 DHFR（SmDHFR）的特性。此外，还使用 X 射线晶体学确定了酶的原子模型，分辨率为 1.95Å。
