植物蛋白质SUMO化修饰体外高效检测系统
收稿日期: 2022-04-12
修回日期: 2022-06-23
网络出版日期: 2022-06-28
基金资助
国家自然科学基金(31871222)
An Effective in Vitro SUMOylation Detection System for Plant Proteins
Received date: 2022-04-12
Revised date: 2022-06-23
Online published: 2022-06-28
蛋白质SUMO化修饰是一种调控蛋白命运的关键修饰方式, 广泛参与植物生长发育及逆境胁迫响应。SUMO化修饰过程主要由激活酶(E1)-结合酶(E2)-连接酶(E3)组成的级联酶促反应催化, 其关键酶组分将SUMO分子缀合至底物蛋白的赖氨酸残基, 形成共价异肽键以完成SUMO化修饰过程。该文报道了1种植物蛋白质SUMO化修饰体外高效检测系统, 通过在大肠杆菌(Escherichia coli)中构建拟南芥(Arabidopsis thaliana) SUMO化修饰的关键通路实现对底物蛋白的SUMO化修饰, 结果可通过免疫印迹进行检测。该系统可以简化植物蛋白质SUMO化修饰的检测流程, 为植物细胞SUMO化修饰的功能研究提供了有力工具。
黄俊文, 冯琦伊, 郑凯勇, 黄俊杰, 王林博, 赖瑞强, 赖建彬, 阳成伟 . 植物蛋白质SUMO化修饰体外高效检测系统[J]. 植物学报, 2022 , 57(4) : 490 -499 . DOI: 10.11983/CBB22080
Protein SUMOylation is a key modification for regulating the fate of proteins and it is widely involved in plant development and stress responses. The SUMO molecules are conjugated to the lysine residues of substrate proteins via isopeptide bonds by enzyme reaction. SUMOylation is mediated by an enzyme cascade composed of a SUMO activating enzyme complex (E1), a SUMO-conjugating enzyme (E2) and usually a SUMO ligase (E3). Here, we report an efficient in vitro detection system for SUMOylation of plant proteins. We established a system for SUMOylation detection of plant proteins by reconstructing the Arabidopsis SUMOylation enzyme cascade in Escherichia coli. Using this system, SUMOylation of several substrates were detected via immunoblotting. Therefore, this system simplifies the SUMOylation detection of plant protein substrates and provides a powerful tool for functional analysis of SUMOylation in plant cells.
Key words： SUMOylation; in vitro; detection system; plant
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