NFAT5 as a transcription factor with an established role in osmotic stress response, has also been revealed to be active under numerous settings, including pathological conditions such as diabetic microvascular complications, chronic arthritis and cancer. Despite these links, current strategies for downregulating NFAT5 activity only relies on indirect modulators, not directly targeting NFAT5, itself. With this study, through using a computational approach, an original peptide was explored to directly target C terminal dimerization of NFAT5 RHR, located in its DNA binding domain. At first, homodimeric NFAT5 RHR bound to its consensus DNA was used for prediction of a preliminary peptide sequence. Possible amino acid replacements for this preliminary peptide were predicted for optimization, which was followed by addition of a cell penetrating peptide sequence. These attempts yielded a small peptide library, which was further investigated for peptide affinities towards C terminal of NFAT5 RHR through molecular docking, 50 ns and 250 ns molecular dynamics simulations, followed by estimation of MM-PBSA based relative binding free energies. Results indicated that after receiving mutations on the preliminary peptide sequence for optimization, a unique peptide could target C terminal dimerization region of NFAT5 RHR through using its cell penetrating peptide sequence. In conclusion, this is the first study presenting computational evidence on identification of a novel peptide capable of directly targeting NFAT5 dimerization. Besides, future implications of these observations were also discussed in terms of methodology and possible applications.

NFAT5 作为一种在渗透应激反应中具有既定作用的转录因子，也已被发现在许多环境下都很活跃，包括糖尿病微血管并发症、慢性关节炎和癌症等病理状况。尽管存在这些联系，但当前下调 NFAT5 活动的策略仅依赖于间接调制器，而不是直接针对 NFAT5 本身。通过这项研究，通过使用一种计算方法，探索了一种原始肽，以直接靶向位于其 DNA 结合域中的 NFAT5 RHR 的 C 末端二聚化。首先，同型二聚体 NFAT5 RHR 与其共有 DNA 结合用于预测初步肽序列。预测该初步肽的可能氨基酸替代品以进行优化，随后添加细胞穿透肽序列。这些尝试产生了一个小肽库，通过分子对接、50 ns 和 250 ns 分子动力学模拟进一步研究了肽对 NFAT5 RHR C 末端的亲和力，然后估计了基于 MM-PBSA 的相对结合自由能。结果表明，在对用于优化的初步肽序列进行突变后，一种独特的肽可以通过其细胞穿透肽序列靶向 NFAT5 RHR 的 C 末端二聚化区域。总之，这是第一项提供关于鉴定能够直接靶向 NFAT5 二聚化的新型肽的计算证据的研究。除了，