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View full-text article in PMC

. 2015 Oct 19;17(9):742–754. doi: 10.1016/j.neo.2015.09.005

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© 2015 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

NPM-ALK is SUMOylated at Lys²⁴ and Lys³² residues. The mutated NPM-ALK recombinant proteins NPM-ALK^K24R and NPM-ALK^K32R were generated using the TnT coupled rabbit reticulocyte kit. In the upper panel, WB was performed to confirm the expression of recombinant NPM-ALK proteins. Wild-type NPM-ALK recombinant protein is present in lane 1, NPM-ALK^K24R recombinant protein in lane 2, and NPM-ALK^K32R recombinant protein in lane 3. In the lower panel, wild-type and mutated recombinant NPM-ALK proteins were subjected to an in vitro SUMOylation assay. SUMOylation was abolished with the K24R mutation for SUMO-1 and SUMO-3 and with the K32R mutation for SUMO-3. The two mutations failed to de-SUMOylate SUMO-2.