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.
bioRxiv : the preprint server for biology
|July 18, 2023
概括
DPc10和calmodulin (CaM) 与泄漏的心脏氨酸受体 (RyR2) 相互作用. DPc10调节了CaM与RyR2的结合,为针对心律失常的药物发现提供了洞察力.
科学领域:
- 心血管研究研究心血管研究
- 分子药理学分子药理学
- 生物物理学的生物物理.
背景情况:
- (Ca2+) 从心肌细胞肉质网膜 (SR) 通过过度活跃的心脏氨酸受体通道 (RyR2) 泄漏是一种亲心律失常的机制.
- 一个外源性,DPc10,检测泄漏的RyR2,而卡尔莫杜林 (CaM) 抑制了RyR2的泄漏,这表明它是一个潜在的治疗标.
- 一个Förster共振能量转移 (FRET) 生物传感器已被设计用于基于这些相互作用的向RyR2的药物发现.
研究的目的:
- 使用FRET阐明在SR囊中DPc10和CaM与RyR2结合之间的相互依赖的分子机制.
- 研究DPc10如何在不同度的Ca2+下影响CaM结合亲和力和RyR2通道活性.
- 了解DPc10与RyR2的合作结合及其通过CaM和FKBP12.6.6的调制.
主要方法:
- 使用FRET与供体-FKBP12.6 (D-FKBP) 和受体-CaM (A-CaM) 来研究CaM/RyR2结合.
- 采用光寿命检测FRET来分析CaM结合动力学和距离.
- 使用受体-DPc10 (A-DPc10) 来研究DPc10/RyR2结合及其通过CaM的调制.
主要成果:
- 在低纳米级Ca2+中,DPc10降低了FRET和CaM/RyR2结合亲和力,降低了CaM位点占用率.
- 在微分子Ca2+中,DPc10降低了FRET,但没有影响CaM/RyR2结合亲和力.
- CaM削弱了A-DPc10/RyR2结合,在微分子比纳米分子Ca2+中更为如此,表明由CaM和FKBP12.6.6调节的合作结合.
结论:
- DPc10以Ca2+依赖的方式调节CaM与RyR2的结合,影响道占用率和距离.
- 在CaM和FKBP12.6的影响下,DPc10与RyR2的合作结合表明了RyR2监管的协调结构变化.
- 这些发现为开发DPc10-CaM范式的治疗策略提供了关键的机制性见解,用于针对RyR2-介导的心律失常的治疗策略.
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