确定TDP-43的N端和RNA结合域之间的相互作用
David D Scott1, Lipsa Jena1,2, Akash Rajaram1
1Department of Pharmacology and Therapeutics, College of Medicine, University of Florida, Gainesville, Florida, USA.
Protein science : a publication of the Protein Society
|September 17, 2025
概括
TAR DNA 结合蛋白 43 (TDP-43) 的 N-终端域 (NTD) 与其 RNA 识别基因 (RRMs) 相互作用,影响 RNA 结合. 这种NTD-RRM交互对于理解TDP-43至关重要.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 神经科学是一个神经科学.
背景情况:
- TAR DNA 结合蛋白 43 千多达尔顿 (TDP-43) 涉及于肌缩性侧面硬化症的发病.
- TDP-43是一种内在无序的蛋白质,具有不同的域:N端域 (NTD),RNA识别动机 (RRMs) 和C端富含甘氨酸的域.
- 之前的研究表明,由于化学探测,TDP-43中发生了性变化,暗示了域间相互作用.
研究的目的:
- 调查TDP-43.3的NTD和RRMs之间的相互作用.
- 确定NTD对TDP-43的RNA结合能力的影响.
- 阐明TDP-43在肌缩侧面硬化症中的作用的结构基础.
主要方法:
- 核磁共振 (NMR) 光谱 (2D [1H,15N] HSQC和Carr-Purcell-Meiboom-Gill) 用于比较TDP-43结构.
- 蛋白质与蛋白质对接以建模NTD-RRM相互作用.
- 微尺度热泳以评估RNA结合亲和力.
主要成果:
- 当NTD存在时,NMR揭示了RRMRNA结合部位的集群转移,即使没有RNA.
- 计算建模提出了NTD对RRM域进行堆叠.
- 实验证据证实了NTD{1-102}和RRMs{102-260}之间的相互作用.
- 与单独RRMs相比,NTD的TDP-43构造显示了显著改变的RNA结合.
结论:
- TDP-43的NTD直接与其RRM域进行交互.
- 这种NTD-RRM相互作用影响了TDP-43结合RNA的能力.
- 了解这些域间相互作用是TDP-43在ALS中的功能和功能障碍的关键.
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