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一个改进的基因编码pH传感器的理性工程基于超临界的pH

Yi Shen¹, Yurong Wen^2,3, Silvia Sposini^4,5

¹Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.

|July 23, 2023

概括

研究人员开发了Lime,这是一款基于超临界pH (SEP) 的新型pH传感器,可提供增强的亮度和灵敏度,用于在内细胞和外细胞形成过程中成像细胞pH的变化.

科学领域:

生物化学生化学
细胞生物学细胞生物学
分子成像学分子成像学

背景情况:

基因编码的pH传感器对于研究细胞过程,如细胞外细胞和细胞内细胞的研究至关重要.
超隐形酸 (SEP) 是一种广泛用于pH成像的绿色光蛋白 (GFP) 变体.
现有的传感器在亮度和灵敏度方面存在局限性.

研究的目的:

理性地设计和开发一种改进的pH敏感光蛋白变体.
与现有的SEP变种相比,提高光亮度和pH敏感度.
描述新变种的结构和应用,命名为Lime.

主要方法:

理性设计原则指导了对SEP的有益突变的引入.
使用X射线晶体学来确定石灰蛋白的结构.
评估了Lime的性能,用于成像细胞事件,如内细胞和外细胞.

主要成果:

石灰呈现出明显更高的光亮度和更高的pH敏感度比SEP.
石灰的晶体结构验证了设计策略和机械学的理解.
石灰在成像内细胞和外细胞的成像中表现出卓越的性能.
石灰及其变体在检测突触释放和神经元电压变化方面显示出实用性.

结论:

石灰代表了基因编码pH传感器的重大进步.
石灰的改进特性使细胞pH动态的成像更有效.
石灰及其变体在神经科学和细胞生物学研究中提供了多方面的应用.

关键词:

光蛋白质是一种光蛋白质.有基因编码的传感器.在 pH 化物中.

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相关概念视频

pH

The potential of hydrogen (pH) is a measure of the acidity or basicity of a water-based solution determined by the concentration of hydronium ions (H3O+). In one liter of pure water at neutral pH, there are 1×10−7 moles of hydronium ions. However, the extensive range of hydronium ion concentrations present in water-based solutions makes measuring pH in moles cumbersome. Therefore, a pH scale was developed to convert moles of hydronium ions into the negative logarithm of the hydronium...