HIGH AFFINITY SUMO TRAPS

摘要

The potential use of these “SUMO receptors” to isolate SUMOylated targets has been considered using the SIM sequences of the SUMO-dependent ubiquitin-protein ligase RNF4. RFN4 contains 4 SIM sequences known to interact with SUMOylated proteins. The capacity of the SIM2 and SIM3 of RNF4 to purify SUMOylated proteins was increased when in the present invention it was disposed in tandem up to 4 SIM sequences. In a preferred embodiment to increase flexibility and functionality of this SUMO-trap, we have changed the natural linkers resulting in a broader capture of SUMOylated proteins. This Tandem SUMO Interacting Motifs (TSIMs) or SUMO-Binding Entities (SUBEs) system disclosed in the present invention is useful to capture polySUMOylated proteins from cell extracts. Therefore, in another embodiment of the present invention, TSIMs or SUBEs can be used for the identification SUMO substrates and the study of SUMO-regulated processes.