相关实验视频

Updated: Aug 20, 2025

Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding

Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding

Published on: June 9, 2017

通过PCS进行纳米体GPS:一种有效的新型G蛋白合受体和其他大型蛋白质的NMR分析方法

Feng-Jie Wu¹, Pascal S Rieder², Layara Akemi Abiko¹

¹Biozentrum, University of Basel, CH-4056Basel, Switzerland.

Journal of the American Chemical Society

|November 17, 2022

概括

这项研究引入了一种新的核磁共振 (NMR) 方法,使用抗体标签来分配蛋白质核. 这种技术有效地绘制了生物宏分子动态,而没有破坏性突变.

科学领域:

生物化学
结构生物学
核磁共振 (NMR) 光谱学

背景情况:

对大型生物大分子 (>1 MDa) 的NMR化学转移变化进行解释需要核特异性赋值.
传统的分配方法,如点突变,是繁的,可以改变蛋白质的功能.

研究的目的:

开发一种高效的NMR方法来分配大型生物大分子中的核.
在没有侵入性基因改造的情况下进行功能动态研究.

主要方法:

使用伪接触转移 (PCS) 核磁共振,使用带有类DOTA化剂标记的抗体片段.
使用抗体结合定位DOTA标签用于大分子核的三角测量.
应用该方法来分配β1-上腺素受体的瓦林和氨酸1H-15N共振.

主要成果:

在β1-上腺素受体中实现了氨酸和氨酸的完整共振分配.
证明了该方法在超过60 Å的距离上三角化核的能力.
在信号传输过程中,通过其电子结构吸收了跨膜螺旋3的显著力量.

结论:

开发的PCS NMR方法为大型生物大分子提供了高效且无破坏性的赋值.
这种技术广泛适用于任何具有可用的结合伙伴的可溶性生物大分子.
能够详细研究复杂生物系统中的功能动态和信号传导机制.

更多相关视频

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy NMR and Microscale Thermophoresis MST

Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy NMR and Microscale Thermophoresis MST

Published on: November 2, 2018

相关概念视频

G-protein Coupled Receptors

G-protein Coupled Receptors

G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.

Applications Of NMR In Biology

Applications Of NMR In Biology

Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.