相关实验视频

Updated: Jun 28, 2025

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale

Published on: March 14, 2019

一个 pentameric KCTD5/CUL3/Gβγ E3 ubiquitin ligase 复合物的结构和动态

Duc Minh Nguyen^1,2, Deanna H Rath¹, Dominic Devost³

¹Princess Margaret Cancer Centre, University Health Network, Toronto, ON M5G 1L7, Canada.

Proceedings of the National Academy of Sciences of the United States of America

|April 16, 2024

概括

KCTD5形成一个多价E3酶复合体,使Gβγ无处不在,调节G蛋白信号传递. 低温电磁探测揭示了无处不在的动态结构和原始化复合体.

科学领域:

生物化学生化学
结构生物学结构生物学
细胞生物学细胞生物学

背景情况:

异构G蛋白是关键的信号分子,通过像无处不在这样的翻译后修饰来调节.
基质受体KCTD5形成了库林-RING E3结合酶复合体 (CRL3^KCTD5) 以使Gβγ亚单元无处不在,从而控制它们的细胞水平.

研究的目的:

阐明CRL3^KCTD5介导的Gβγ的无处不在的结构和机制基础.
调查KCTD5寡合化在形成多价E3结合酶复合物的作用.

主要方法:

电子显微镜 (cryo-EM) 用于确定 KCTD5/CUL3^NTD/Gβ₁γ₂组件的结构.
扩展复合物的结构分析,包括全长的CUL3,RBX1和ARIH1~ubiquitin结合物.

主要成果:

冷-EM结构显示了一个动态的5:5:5 KCTD5/CUL3/NTD/Gβγ复合体,具有显著的域间旋转.
CRL3^KCTD5与ARIH1形成了一个E3-E3超组件,用于Gβγ无处不在.
结构数据表明,在ARIH1~ubiquitin被定位为高效的Gβ无处不在的基础复合体.

结论:

基质受体的寡合化,正如KCTD5所示,可以产生多价E3结合酶复合体.
KCTD5的内在动力学有助于在CRL3复合体内定位结构化的Gβγ点,使其无处不在.

关键词:

这种BTB蛋白质是BTB蛋白质.G 蛋白质是 G 蛋白质的组成部分.化电子显微镜技术的电子显微镜无处不在的合酶

更多相关视频

In Vitro Analysis of E3 Ubiquitin Ligase Function

In Vitro Analysis of E3 Ubiquitin Ligase Function

Published on: May 14, 2021

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

相关概念视频

Covalently Linked Protein Regulators

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....

The Proteasome Structure

The Proteasome Structure

The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
The proteasome is an...

Protein Complex Assembly

Protein Complex Assembly

Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types. Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...

Protein Complexes with Interchangeable Parts

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...

Regulated Protein Degradation

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...

The Proteasome

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...