This chapter deals with the fluorescence detection of SUMOylation and deSUMOylation in semi-intact cultured human cells, the so-called “in situ SUMOylation assay” and the “in situ deSUMOylation assay,” respectively. In the in situ SUMOylation assay, the recombinant green-fluorescence protein fused to the SUMO1 (GFP-SUMO1) protein is used to visualize the nuclear rim, nucleolus, and nuclear bodies. These GFP signals represent cellular regions where SUMOylation efficiently takes place. If the recombinant SUMOspecific protease SENP1-catalytic domain is added after in situ SUMOylation, GFP signals can be erased. Therefore, the in situ SUMOylation assay can be used to assess deSUMOylation enzymatic activity.
CITATION STYLE
Yuasa, E., & Saitoh, H. (2016). In situ SUMoylation and DeSUMoylation assays: Fluorescent methods to visualize sumoylation and DeSUMoylation in permeabilized cells. In Methods in Molecular Biology (Vol. 1475, pp. 151–159). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6358-4_11
Mendeley helps you to discover research relevant for your work.