对心脏瑞诺丁受体FRET生物传感器的分子机制 病理性泄漏状态
Bengt Svensson1, Florentin R Nitu1, Robyn T Rebbeck1
1Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
International journal of molecular sciences
|August 26, 2023
概括
一种外源性 (DPc10) 和calmodulin (CaM) 调节心脏氨酸受体2 (RyR2) 通道活性. 了解它们的相互作用揭示了针对RyR2在心律失常的药物发现中的机制.
科学领域:
- 心脏病学 心脏病学
- 分子生物学分子生物学
- 生物物理学的生物物理.
背景情况:
- 通过心肌细胞中的过度活跃的心脏氨酸受体通道 (RyR2) 泄漏的Ca2+有助于心律失常.
- 一种外源性 (DPc10) 促进了RyR2的泄漏,而卡尔莫杜林 (CaM) 则抑制了它,这表明它是一个潜在的治疗标.
研究的目的:
- 阐明DPc10与CaM结合RyR2.2之间的相互依赖的分子机制.
- 为开发FRET生物传感器提供信息,用于针对RyR2.2的药物发现.
主要方法:
- 福斯特共振能量转移 (FRET) 生物传感器利用供体-FKBP12.6 (D-FKBP) 和受体-CaM (A-CaM) 来研究RyR2-CaM相互作用.
- 光寿命检测FRET用于分析Ca2+依赖性和结合动力学.
- 使用受体-DPc10 (A-DPc10) 来研究DPc10/RyR2结合和合作性.
主要成果:
- 在纳米Ca2+中,DPc10降低了CaM/RyR2结合亲和力和FRETmax,但仅在微分子Ca2+中降低了FRETmax.
- DPc10降低了纳米分子Ca2+中的RyR2 CaM结合位点占用率,并增加了D-FKBP/A-CaM距离.
- CaM削弱了DPc10/RyR2结合,在微分子Ca2+中具有更大的效果;结合是合作性的,并且依赖于CaM和FKBP.
结论:
- 在调节RyR2.2,DPc10和CaM表现出Ca2+依赖的相互依赖性.
- 这些发现为DPc10-CaM范式提供了机制性的见解,用于开发针对RyR2功能障碍的治疗方法.
- 合作结合表明由这些调节器调节的RyR2原体的协调结构变化.
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