Department of Biochemistry and Biophysics, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, USA.
Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, USA.
Biomol NMR Assign. 2021 Apr;15(1):197-202. doi: 10.1007/s12104-021-10006-x. Epub 2021 Jan 24.
Human thymidylate synthase (hTS) is a 72 kDa homodimeric enzyme responsible for the conversion of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP), making it the sole source of de novo dTMP in human cells. As a result, hTS is an attractive anti-cancer therapeutic target. Additionally, hTS is known to possess a number of interesting biophysical features, including adoption of active and inactive conformations, positively cooperative substrate binding, half-the-sites activity, and interacting with its own mRNA. The physical mechanisms underlying these properties, and how they may be leveraged to guide therapeutic development, are yet to be fully explored. Here, as a preface to detailed NMR characterization, we present backbone amide and ILVM methyl resonance assignments for hTS in apo and dUMP bound forms. In addition, we present backbone amide resonance assignments for hTS bound to a substrate analog and the native cofactor.
人胸苷酸合成酶（hTS）是一种 72kDa 的同二聚体酶，负责将脱氧尿苷单磷酸（dUMP）转化为脱氧胸苷单磷酸（dTMP），使其成为人细胞中从头合成 dTMP 的唯一来源。因此，hTS 是一个有吸引力的抗癌治疗靶点。此外，hTS 已知具有许多有趣的生物物理特性，包括采用活性和非活性构象、正协同底物结合、半位活性以及与自身 mRNA 相互作用。这些特性的物理机制，以及如何利用这些机制来指导治疗药物的开发，仍有待充分探索。在这里，作为详细 NMR 特征描述的前言，我们展示了 apo 和 dUMP 结合形式的 hTS 的酰胺基和 ILVM 甲基共振分配。此外，我们还展示了 hTS 与底物类似物和天然辅因子结合的酰胺基共振分配。
