Chinese Bulletin of Botany ›› 2022, Vol. 57 ›› Issue (4): 490-499.DOI: 10.11983/CBB22080

• INVITED PROTOCOL • Previous Articles     Next Articles

An Effective in Vitro SUMOylation Detection System for Plant Proteins

Huang Junwen, Feng Qiyi, Zheng Kaiyong, Huang Junjie, Wang Linbo, Lai Jianbin Lai Ruiqiang, Yang Chengwei   

  1. Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou 510631, China
  • Received:2022-04-12 Revised:2022-06-23 Online:2022-07-01 Published:2022-07-14
  • About author:First author contact: These authors contributed equally to this paper

Abstract: 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