The goal of this manuscript is to present an outline for the comprehensive biochemical and functional studies of the RING-type E3 ubiquitin ligases. This multistep pipeline, with detailed protocols, validates an enzymatic activity of the tested protein and demonstrates how to link the activity to function.
Kud, J., Wang, W., Yuan, Y., Caplan, A., Kuhl, J. C., Dandurand, L., Xiao, F. Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta. J. Vis. Exp. (154), e60533, doi:10.3791/60533 (2019).