相关实验视频
Updated: Jun 30, 2026
12:30
Monitoring ER/SR Calcium Release with the Targeted Ca2+ Sensor CatchER+
Published on: May 19, 2017
基于绿色光蛋白和calmodulin的Ca2+光指标
1Department of Pharmacology, University of California, San Diego, La Jolla 92093-0647, USA.
Nature
|August 28, 1997
概括
研究人员开发了一种基因编码的光 (Ca2+) 指示器,称为驼. 这些驼使得可视化局部Ca2+信号在有机细胞和活细胞中的细胞质.
科学领域:
- 细胞生物学 细胞生物学
- 生物化学 生物化学
- 分子生物学分子生物学
背景情况:
- 局部 (Ca2+) 信号对于细胞过程至关重要,但很难测量.
- 现有的方法难以捕捉细胞内Ca2+的空间和时间动态.
研究的目的:
- 开发新的,基因编码的光指标,用于测量局部Ca2+信号.
- 为了可视化Ca2+动态在特定的亚细胞区间,如细胞质,核和内分泌网膜.
主要方法:
- "驼"的构建,它们是光蛋白 (GFP变体),calmodulin和M13的协同融合.
- 利用GFP变体之间的光共振能量转移 (FRET) 来检测与calmodulin结合的Ca2+.
- 在活着的HeLa细胞中对驼进行基因编码,以便在活着的HeLa细胞中有针对性的表达和可视化.
主要成果:
- 驼成功地可视化了细胞质,细胞核和内 плазма网膜中的局部Ca2+动态.
- 静止Ca2+度在内质网膜中测量在60-400微米之间.
- 活化Ca2+度在内质网膜中范围为1-50微米.
- 此外,FRET还展示了监测蛋白质异构化的能力.
结论:
- 驼是测量局部Ca2+信号和活细胞中蛋白质相互作用的有效工具.
- 开发的指标使得可以详细研究Ca2+在各种亚细胞区的信号传递.
- 这项技术推动了对细胞Ca2+动态和功能的理解.
相关概念视频
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
01:16Calmodulin-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,...
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...