协议确定Shigella sonnei中核酸第二信使的信号网络
Binbin Cui1, Xiayu Chen2, Mingfang Wang2
1Pharmacy Department, The Affiliated LiHuiLi Hospital of Ningbo University, Ningbo 315046, China; School of Pharmaceutical Sciences (Shenzhen), Shenzhen Campus of Sun Yat-sen University, Sun Yat-sen University, Shenzhen 518107, China.
STAR protocols
|February 11, 2026
概括
本研究详细介绍了一种检测Shigella sonnei.中的循环AMP (cAMP) 和循环二GMP (c-di-GMP) 信号网络的协议. 这些方法可以对这些关键的细菌第二信使系统进行全面分析.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 细菌的第二信使信号系统,包括循环AMP (cAMP) 和循环di-GMP (c-di-GMP),在调节各种细胞过程中起着至关重要的作用.
- 了解这些信号网络对于破译细菌生理学和病变产生至关重要.
- 希格拉·索内伊 (Shigella sonnei) 是一个重要的人类病原体,其生存和毒性依赖于复杂的信号通路.
研究的目的:
- 提出一个全面的协议,用于检测和分析cAMP和c-di-GMP信号网络在Shigella sonnei.
- 为研究细菌第二信使系统的研究人员提供标准化的方法.
主要方法:
- 微尺度热泳 (MST) 用于结合分析.
- 定量逆转录PCR (RT-qPCR) 用于基因表达分析.
- 超高性能液态染色体质谱仪 (LC-MS) 用于代谢物量化.
- 电泳运动转移试验 (EMSA) 用于DNA-蛋白相互作用.
- 细菌腺酸环酶双混合体 (BACTH) 测定蛋白质-蛋白质相互作用.
- 分子对接用于计算分析.
主要成果:
- 本协议成功地整合了多种技术,使cAMP和c-di-GMP信号网络组件的检测成为可能.
- 这项研究提供了一个框架,用于描述Shigella sonnei中的这两个关键的第二信使系统之间的相互作用.
- 使用该协议可以阐明信号网络中的特定分子相互作用和调节机制.
结论:
- 开发的协议为研究细菌第二信使信号网络提供了一种强大而通用的方法.
- 这种方法有助于更深入地了解控制Shigella sonnei病变的分子机制.
- 该协议可以适应在其他细菌物种中研究类似的信号系统.
相关概念视频
01:15Amplifying Signals via Second Messengers
8.8K
Many receptor binding ligands are hydrophilic; they do not cross the cell membrane but bind to cell-surface receptors. Thus, their message must be relayed by second messengers present in the cell cytoplasm. There are several second messenger pathways, each with its own way of relaying information. For example, the G protein-coupled receptors can activate both phosphoinositol and cyclic AMP (cAMP) second messenger pathways. The phosphoinositol pathway is active when the receptor induces...
8.8K
01:12What are Second Messengers?
90.8K
Because many receptor binding ligands are hydrophilic, they do not cross the cell membrane and thus their message must be relayed to a second messenger on the inside. There are several second messenger pathways, each with their own way of relaying information. G-protein coupled receptors can activate both phosphoinositol and cyclic AMP (cAMP) second messenger pathways. The phosphoinositol path is active when the receptor induces phospholipase C to hydrolyze the phospholipid,...
90.8K
01:08Nucleotide Excision Repair
40.9K
Overview
40.9K
01:21Cell-surface Signaling
54.7K
Hormones—or any molecule that binds to a receptor, known as a ligand—that are lipid-insoluble (water-soluble) are not able to diffuse across the cell membrane. In order to be able to affect a cell without entering it, these hormones bind to receptors on the cell membrane. When a first messenger, a hormone, binds to a receptor, a signal cascade is set off, causing second messengers, proteins inside the cell, to become activated, resulting in downstream effects.
54.7K
02:26Protein Networks
4.6K
An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
4.6K
01:30Bacterial Signaling
41.3K
Bacterial signaling can occur within bacteria (intracellular) or between bacteria (intercellular). At times, a group of bacteria behaves like a community. To achieve this, they engage in quorum sensing, the perception of higher cell density that causes changes in gene expression. Quorum sensing involves both extracellular and intracellular signaling. The signaling cascade starts with a molecule called an autoinducer (AI). Individual bacteria produce AIs that move out of the bacterial cell...
41.3K