相关实验视频
Updated: Jul 13, 2026

09:16
Oligopeptide Competition Assay for Phosphorylation Site Determination
Published on: May 18, 2017
循环-AMP-响应交换因子Epac2在其自抑制状态中的结构
Holger Rehmann1, Joost Das, Puck Knipscheer
1Department of Physiological Chemistry and Centre for Biomedical Genetics, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands.
Nature
|February 3, 2006
概括
由循环AMP (Epac) 直接激活的交换蛋白调节细胞过程. 该研究揭示了Epac2的自身抑制结构,详细介绍了其调节和催化区域以及cAMP介导激活的机制.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 细胞信号传递 细胞信号传递
背景情况:
- Epac 蛋白质是 Rap1 和 Rap2 GTPases 的关氨酸核酸交换因子 (GEF).
- 它们由循环AMP (cAMP) 直接调节,并参与细胞粘附和胰岛素分泌.
研究的目的:
- 要确定全长 Epac2.2 的三维结构.
- 阐明自抑制状态和cAMP诱导激活的机制.
主要方法:
- 使用X射线晶体学来解决Epac2.2的结构.
- 进行突变分析以调查激活机制.
主要成果:
- 确定了全长Epac2在自身抑制状态中的结构.
- 一个"开关板"结构和"离子锁"被确定为关键的抑制元素.
- 提出了一个cAMP诱导的激活模型,涉及硬的身体运动.
结论:
- 结构揭示了Epac2在没有cAMP的情况下是如何自我抑制的.
- 这些发现提供了关于cAMP调节蛋白激活的普遍机制的见解.
相关概念视频
Amplifying Signals via Enzymatic Cascade
When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze the...
GPCR Desensitization
G protein-coupled receptor (GPCR) signaling plays a crucial role in cell functioning. GPCR desensitization is an equally essential process. It allows cells to respond to changing environments and regain sensitivity to new stimuli while preventing unnecessary stimulation when no longer needed. Prolonged exposure to stimuli leads to GPCR desensitization. It involves blocking the receptors from binding and activating additional G proteins. This inhibits activation of downstream effectors, thereby...
GPCRs Regulate Adenylyl Cylase Activity
Some GPCRs transmit signals through adenylyl cyclase (AC), a transmembrane enzyme. AC helps synthesize second messenger cyclic adenosine monophosphate (cAMP). AC catalyzes cyclization reaction and converts ATP to cAMP by releasing a pyrophosphate. The pyrophosphate is further hydrolyzed to phosphate by the enzyme pyrophosphatase, which drives cAMP synthesis to completion. However, cAMP is rapidly degraded to 5′ AMP by the enzymes phosphodiesterase (PDE), preventing overstimulation of cells.
Two...
Two...
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,...
Global Regulatory Systems
Global regulatory systems in bacteria enable rapid and coordinated responses to environmental changes by integrating sensory inputs with gene expression, ensuring efficient adaptation to fluctuating conditions. Key global regulatory mechanisms include regulons, two-component systems, sigma factors, and secondary messengers.Regulons and Global RegulatorsA regulon is a collection of genes and operons controlled by a common global regulator. These regulators enable bacteria to prioritize resource...

