相关实验视频

Updated: Jul 10, 2025

TurboID-Based Proximity Labeling for In Planta Identification of Protein-Protein Interaction Networks

TurboID-Based Proximity Labeling for In Planta Identification of Protein-Protein Interaction Networks

Published on: May 17, 2020

基于TurboID的近距离标记:一种解读植物中蛋白质之间的相互作用的方法

Yuanyuan Li¹, Yongliang Zhang², Savithramma P Dinesh-Kumar³

¹Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, USA.

Methods in molecular biology (Clifton, N.J.)

|November 21, 2023

概括

TurboID近距离标记 (PL) 为研究植物蛋白与蛋白相互作用 (PPI) 提供了一个更快,室温的方法. 这种优化的协议克服了旧技术的局限性,使植物细胞过程的有效分析成为可能.

科学领域:

植物分子生物学植物分子生物学
生物化学生化学
蛋白质组学是指蛋白质组学.

背景情况:

蛋白与蛋白相互作用 (PPI) 对细胞过程至关重要.
传统的PPI分析方法在植物系统中存在局限性,包括毒性和长潜伏时间.
酶催化近距离标记 (PL) 技术提供了替代方案,但需要对植物进行优化.

研究的目的:

描述一种基于TurboID的近距离标签 (PL) 的优化协议,用于研究PPI的工厂.
为了证明TurboID-PL与BioID相比在标记内源植物蛋白中的有效性.
为研究各种植物物种的PPI网络提供一种可适应的方法.

主要方法:

基于农细菌的蛋白质的短暂表达.
生物素处理和随后的蛋白质提取.
免费生物的去除,定量,以及生物化蛋白的亲和力净化.
使用TurboID-PL与植物病毒免疫受体 (NLR类) 作为模型系统.

主要成果:

基于TurboID的PL在标记植物内源蛋白质方面表现优于BioID.
优化的协议可以在室温下快速贴近标签.
成功应用了涉及植物免疫受体的PPI研究方法.

结论:

基于TurboID的PL是研究植物PPI的高效和多功能工具.
描述的协议克服了以前的局限性,促进了植物分子生物学研究.
这种方法可以很容易地适应各种植物物种和PPI网络分析.

关键词:

生物化生物化在NLR免疫受体中,NLR免疫受体是最重要的.蛋白质蛋白质相互作用靠近标签的标签.这是一个TurboIDID.

更多相关视频

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants

Published on: September 16, 2022

Protein-protein Interactions Visualized by Bimolecular Fluorescence Complementation in Tobacco Protoplasts and Leaves

Protein-protein Interactions Visualized by Bimolecular Fluorescence Complementation in Tobacco Protoplasts and Leaves

Published on: March 9, 2014

相关概念视频

Protein-protein Interfaces

Protein-protein Interfaces

Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a...

Protein Networks

Protein Networks

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,...

Cell Signaling in Plants

Cell Signaling in Plants

Plant cells communicate to coordinate their cycle of growth, flowering and fruiting, and activities in roots, shoots, and leaves in response to the changing environmental conditions. Plant signaling is distinct from animal signaling. Plants primarily utilize enzyme-linked receptors, whereas the largest class of cell-surface receptors in animals are G-protein coupled receptors (GPCRs). Unlike animals, receptor tyrosine kinases are rare in plants. Instead, plants have a diverse class of...