Additional file 3: of Wheat germ-based protein libraries for the functional characterisation of the Arabidopsis E2 ubiquitin conjugating enzymes and the RING-type E3 ubiquitin ligase enzymes

Summary of Arabidopsis RING clones used in this study. We demonstrate their expression and their ubiquitination activities. Summary of Arabidopsis RING proteins used in this study, their expression in a wheat germ cell-free system and their ubiquitination activities. ‘M. wt. (Da)’ indicates the expected molecular weight of the expressed proteins according to RAFL database. ‘High molecular smear’ indicates the detection of smear: awhen RING protein expression was detected by immunoblot analysis or bwhen analysed in ubiquitination assays without the addition of E1 or E2. ‘Activity with UBC8/10’ indicates the E3 ligase activity of selected RING proteins tested with cAtUBC8 [14] or dAtUBC10 in this study. ‘Activity with other E2s’ indicates the E3 ligase activity of selected RING proteins tested with evarious E2s [13] or fin this study. ‘Truncated’ refers to protein that migrated more than 20 KDa less than the expected size. ‘Low expression’ refers to proteins expressed at relatively low levels. Abbreviations: yes, detected; No, not detected; nd, not tested. (XLSX 69 kb)