Abstract
Ubiquitination of proteins is one of post-translational modification in eukaryote and is catalyzed by the three components, E1, E2 and E3. Although main function of the ubiquitination is known to lead to protein degradation, more recent studies showed that ubiquitination might function as a regulator of signal transduction and gene expression. Genomic sequence of model plant Arabidopsis thaliana revealed more than 500 genes encoding the components of ubiquitination. Thus plant might have a highly complicated ubiquitination network. In this study, by combining the wheat cell-free expression system and high-throughput detection system, AlphaScreen, we aimed to construct the comprehensive method for analysis of the ubiquitination network. Twenty-four of 32 E2 genes from RAFL were expressed as soluble proteins, and twenty of them were markedly detected the ubiquitination from rabbit E1. Now we are challenging to investigate the ubiquitination of proteins involved in signal transduction by using this method.
