一种光蛋白C端核聚变敲进与TRPA1是功能性的,但不是TRPC5的
Aaron Tragl1, Alexandra Ptakova2, Viktor Sinica3
1Friedrich-Alexander-Universität Erlangen-Nürnberg, Department of Anesthesiology, Krankenhausstraße 12, 91054 Erlangen, Germany.
Biochimica et biophysica acta. Molecular cell research
|December 11, 2024
概括
研究人员为TRPC5和TRPA1通道开发了新的光蛋白记者小鼠. 虽然TRPA1记者显示光,但两个通道都经历了功能障碍,突出显示了三神经元中C终端的重要性.
科学领域:
- 分子生物学分子生物学
- 神经科学是一个神经科学.
- 遗传学 是一个遗传学.
背景情况:
- 光蛋白 (FP) 记者是体内成像的宝贵工具.
- 暂时受体潜力 (TRP) 通道在感官生理学中起着至关重要的作用.
- 了解TRP通道功能需要精确的细胞和组织水平的研究.
研究的目的:
- 使用FP C端融合生成TRPC5和TRPA1报告者小鼠.
- 为研究TRP通道生理学和相互作用提供改进的模型.
- 研究FP标记在TRPC5和TRPA1通道上的功能后果.
主要方法:
- 产生的转基因结构具有C端FP融合 (mCherry为TRPC5,mTagBFP为TRPA1),由3*GGGGS序列连接在一起.
- 通过致卵子微注射创建了记者小鼠.
- 分析了记者小鼠使用双光子显微镜,免疫组织化学和成像的三腺.
主要成果:
- 成功整合了TRPC5-mCherry和TRPA1-mTagBFP结构的DNA和RNA.
- TRPC5-mCherry小鼠在蛋白质水平上没有显示可检测的mCherry光.
- TRPA1-mTagBFP记者小鼠在感觉神经元中表现出mTagBFP光,但TRPA1通道功能受损.
结论:
- 对TRP通道的C端FP标记需要对特定的通道子类型进行仔细的构造设计和验证.
- 远端C端似乎对三神经元中TRPC5和TRPA1通道的功能至关重要.
- 在TRPA1和TRPC5记者小鼠中观察到的功能障碍需要进一步研究C终端角色.
更多相关视频
10:41Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
Published on: February 17, 2017
8.0K
14:02Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
Published on: April 9, 2018
8.4K
相关概念视频
Transfer RNA Synthesis
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Nuclear Protein Sorting
Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
Vesicular Tubular Clusters
After budding out from the ER membrane, some COPII vesicles lose their coat and fuse with one another to form larger vesicles and interconnected tubules called vesicular tubular clusters or VTCs. These clusters constitute a compartment at the ER-Golgi interface known as ERGIC (Endoplasmic Reticulum Golgi Intermediate Compartment). The ERGIC is a mobile membrane-bound cargo transport system that sorts proteins secreted from ER and delivers them to the Golgi.
With the help of motor proteins such...
With the help of motor proteins such...
Cotranslational Protein Translocation
Translocation of proteins across membranes is an ancient process that occurs even in bacteria and archaebacteria. In fact, the components of the translocation machinery are still conserved between prokaryotes and eukaryotes.
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
Tail-anchoring of Proteins in the ER Membrane
Tail-anchored, or TA, proteins are estimated to make up to 3-5% of membrane proteins found in the eukaryotic cell. Such proteins have a single transmembrane domain located approximately 30 amino acid residues upstream from the C-terminal end. As a result, the signal recognition particle (SRP) cannot guide a TA protein to the ER membrane for cotranslational insertion. Hence, they are integrated into the ER membrane post-translationally using their C-terminal end as the anchor. TA proteins...
Catenins
Catenins are characterized by multiple binding domains and dynamic structures that allow them to function as linker proteins in cell junction complexes. All catenins, except α-catenin, contain a characteristic protein sequence called the armadillo repeat and are therefore also called armadillo proteins.
Catenins in Cell Junctions
Catenins bind to cell adhesion molecules such as cadherins and link them to different cytoskeletal proteins depending on the type of cell junction. At the adherens...
Catenins in Cell Junctions
Catenins bind to cell adhesion molecules such as cadherins and link them to different cytoskeletal proteins depending on the type of cell junction. At the adherens...
