相关实验视频
Updated: Jul 27, 2026

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Detecting Anastasis In Vivo by CaspaseTracker Biosensor
Published on: February 1, 2018
在细胞中成像蛋白激酶卡尔法激活
1Protein Phosphorylation Laboratory and Cell Biophysics Laboratory, Imperial Cancer Research Fund (ICRF), 44 Lincoln's Inn Fields, London, WC2A 3PX, UK.
概括
这项研究确定了一种新的方法来跟踪蛋白激酶卡尔法 (PKCalpha) 激活,使用光共振能量转移 (FRET) 和显微镜. 这种方法可实时可视化细胞和组织中的酶活性.
科学领域:
- 生物物理学的生物物理.
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 光共振能量转移 (FRET) 和光终身成像显微镜 (FLIM) 为研究蛋白质功能提供了强大的工具.
- 具有空间分辨率的FRET/FLIM可以监测活体和固定生物样本中的蛋白质活性.
研究的目的:
- 开发和验证一种基于FRET/FLIM的方法,用于动态成像蛋白激酶卡尔法 (PKCalpha) 激活.
- 证明这种方法在活细胞,固定细胞和病变组织中的实用性.
主要方法:
- 使用FLIM测量的FRET来检测PKCalpha激活.
- 采用光标记的酸化部位特异性抗体来识别激活的PKCalpha.
- 将该技术应用于活细胞和固定培养细胞,以及病理样本.
主要成果:
- 成功识别并利用PKCalpha激活的动态标记物.
- 启用了在细胞环境中PKCalpha激活的实时成像.
- 对实验性细胞培养和临床病理样本均有证明的适用性.
结论:
- 空间分辨率的FRET/FLIM是一种可行的方法,用于追踪蛋白质的催化活性和功能状态.
- 这种方法提供了PKCalpha激活的动态读数.
- 该方法在生物和临床研究中具有广泛的潜力.
相关概念视频
Phosphorylation
The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
Phosphorylation
The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
Protein Kinases and Phosphatases
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein Kinases and Phosphatases
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
cAMP-dependent Protein Kinase Pathways
Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
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,...
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...

