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Updated: Sep 14, 2025

Pull-down of Calmodulin-binding Proteins

Pull-down of Calmodulin-binding Proteins

Published on: January 23, 2012

旋转顶形纳米探针用于直接可视化单个calmodulin分子中的合作Ca2+-结合诱导的合规切换

Rui Liu¹, Zhuodong Tang¹, Qing Xia¹

¹State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, 163 Xianlin Avenue, Nanjing 210023, China.

Journal of the American Chemical Society

|July 22, 2025

概括

研究人员开发了一种新方法,使用纳米级旋转顶观察calmodulin.

科学领域:

生物物理学的生物物理.
单分子生物物理学的单分子生物物理.
蛋白质动力学蛋白质动力学

背景情况:

了解calmodulin的结构动态对于信号传递至关重要.
现有的单分子技术对于快速动态的时间分辨率有限.
卡尔莫杜林 (CaM) 在结合时经历了显著的形状变化.

研究的目的:

开发一种高速的,以毫秒分辨率的方法来研究单个卡尔莫杜林的动态.
在calmodulin分子中直接解决特定域的构造变化.
在单个分子水平上量化-CaM相互作用和结合动力学.

主要方法:

使用高速纳米级旋转顶 (NST) 作为纳米探测器.
在金纳米基上构建了一个单分子CaM突起.
测量的随机,热驱动的旋转调节的CaM形态状态.

主要成果:

在apo-CaM (性) 和Ca2+结合的CaM (性) 状态中观察到明显的旋转特征.
实现了对CaM形态切换的毫秒分辨率检测.
动力分析显示,两个Ca2+离子与C端域的合作结合 (希尔系数~1.81).

结论:

该NST平台能够直接,高时间分辨率观察单个CaM的形状动态.
这种方法提供了对蛋白质 - 连接体相互作用和形状动力学的定量见解.
提供了对介导信号和动态蛋白质功能的新理解.

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相关概念视频

Calmodulin-dependent Signaling

Calmodulin-dependent Signaling

Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...