当前位置: X-MOL 学术J. Biol. Chem. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Cyclic peptides with a distinct arginine-fork motif recognize the HIV trans-activation response RNA in vitro and in cells.
Journal of Biological Chemistry ( IF 5.5 ) Pub Date : 2021-11-09 , DOI: 10.1016/j.jbc.2021.101390
Sai Shashank Chavali 1 , Sachitanand M Mali 2 , Rachel Bonn 1 , Abhijith Saseendran Anitha 2 , Ryan P Bennett 3 , Harold C Smith 4 , Rudi Fasan 2 , Joseph E Wedekind 1
Affiliation  

RNA represents a potential target for new antiviral therapies, which are urgently needed to address public health threats such as the human immunodeficiency virus (HIV). We showed previously that the interaction between the viral Tat protein and the HIV-1 trans-activation response (TAR) RNA was blocked by TB-CP-6.9a. This cyclic peptide was derived from a TAR-binding loop that emerged during lab evolution of a TAR-binding protein (TBP) family. Here we synthesized and characterized a next-generation, cyclic-peptide library based on the TBP scaffold. We sought to identify conserved RNA-binding interactions and the influence of cyclization linkers on RNA binding and antiviral activity. A diverse group of cyclization linkers, encompassing disulfide bonds to bicyclic aromatic staples, was used to restrain the cyclic peptide geometry. Thermodynamic profiling revealed specific arginine-rich sequences with low to submicromolar affinity driven by enthalpic and entropic contributions. The best compounds exhibited no appreciable off-target binding to related molecules, such as BIV TAR and human 7SK RNAs. A specific arginine-to-lysine change in the highest affinity cyclic peptide reduced TAR binding by tenfold, suggesting that TBP-derived cyclic peptides use an arginine-fork motif to recognize the TAR major groove while differentiating the mode of binding from other TAR-targeting molecules. Finally, we showed that HIV infectivity in cell culture was reduced in the presence of cyclic peptides constrained by methylene or naphthalene-based linkers. Our findings provide insight into the molecular determinants required for HIV-1 TAR recognition and antiviral activity. These findings are broadly relevant to the development of antivirals that target RNA molecules.

中文翻译:

具有独特精氨酸叉基序的环状肽可在体外和细胞中识别 HIV 反式激活反应 RNA。

RNA 代表了新的抗病毒疗法的潜在目标,迫切需要解决人类免疫缺陷病毒 (HIV) 等公共卫生威胁。我们之前表明,病毒 Tat 蛋白和 HIV-1 反式激活反应 (TAR) RNA 之间的相互作用被 TB-CP-6.9a 阻断。这种环肽来源于在 TAR 结合蛋白 (TBP) 家族的实验室进化过程中出现的 TAR 结合环。在这里,我们合成并表征了基于 TBP 支架的下一代环状肽库。我们试图确定保守的 RNA 结合相互作用以及环化接头对 RNA 结合和抗病毒活性的影响。一组不同的环化接头,包括与双环芳香主食的二硫键,用于限制环肽的几何形状。热力学分析揭示了由焓和熵贡献驱动的具有低至亚微摩尔亲和力的特定富含精氨酸的序列。最好的化合物与相关分子(如 BIV TAR 和人类 7SK RNA)没有明显的脱靶结合。最高亲和力环肽中精氨酸到赖氨酸的特定变化将 TAR 结合减少了十倍,这表明 TBP 衍生的环肽使用精氨酸叉基序来识别 TAR 大沟,同时将结合模式与其他 TAR 靶向区分开来分子。最后,我们发现细胞培养物中的 HIV 感染性在存在受亚甲基或萘基接头约束的环肽的情况下降低。我们的研究结果提供了对 HIV-1 TAR 识别和抗病毒活性所需的分子决定因素的深入了解。
更新日期:2021-11-09
down
wechat
bug