暴露隐藏的高 afinity RNA 构成状态
Nicole I Orlovsky1, Hashim M Al-Hashimi1,2, Terrence G Oas1,2
1Department of Biochemistry , Duke University Medical Center , Durham , North Carolina 27710 , United States.
Journal of the American Chemical Society
|December 10, 2019
概括
了解RNA结合亲和力需要考虑构造性惩罚. 这项研究表明,即使是整体弱的RNA-连接体结合,也可能涉及到对特定的预折叠RNA状态的强结合,从而影响药物发现.
科学领域:
- 生物化学
- 结构生物学
- 分子生物物理学
背景情况:
- RNA结合亲和力受形态变化和相关的能量成本的影响,称为"形态惩罚".
- 测量这些惩罚是具有挑战性的,因为结合的RNA结构通常在未结合状态下具有不可检测的群体.
研究的目的:
- 为结合的RNA结构变化和连接物结合开发热力学模型.
- 在HIV-1 TAR RNA-阿尔金胺 (ARG) 相互作用模型系统中量化构造性处罚.
主要方法:
- 使用核磁共振 (NMR) 化学转移扰动和放松分散.
- 应用贝叶斯推理对12个状态模型来估计低人口状态的能量.
- 研究了HIV-1 TAR RNA与阿尔基尼胺 (ARG) 作为Tat蛋白模仿物的相互作用.
主要成果:
- 根据 TAR 形状和 ARG 占用率,确定了至少四种不同的 RNA 中介物.
- 发现具有两个ARG连接体的主导绑定 TAR 构造具有未绑定的低于检测极限平衡群体.
- 证明ARG结合了预折叠的TAR形状,具有纳米分子亲和力,尽管整体亲和力明显较弱 (0.2mM).
结论:
- 符合性处罚显著决定了RNA-连接体结合性,并可以推动结合性合作性.
- 准确预测RNA识别需要考虑构造转换的能量成本.
- 这些发现对RNA向药物发现和RNA蛋白相互作用的理解有意义.
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