biostudies-literatureSchou JNovo Nordisk Foundation Center for Protein Researche100692https://www.ebi.ac.uk/biostudies/studies/S-EPMC4074068biostudies-literatureUnknown9(6)During mitosis large alterations in cellular structures occur rapidly, which to a large extent is regulated by post-translational modification of proteins. Modification of proteins with the small ubiquitin-related protein SUMO2/3 regulates mitotic progression, but few mitotic targets have been identified so far. To deepen our understanding of SUMO2/3 during this window of the cell cycle, we undertook a comprehensive proteomic characterization of SUMO2/3 modified proteins in mitosis and upon mitotic exit. We developed an efficient tandem affinity purification strategy of SUMO2/3 modified proteins from mitotic cells. Combining this purification strategy with cell synchronization procedures and quantitative mass spectrometry allowed for the mapping of numerous novel targets and their dynamicsPloS oneComprehensive identification of SUMO2/3 targets and their dynamics during mitosis.PMC4074068PI Jakob NilssonPI Jesper Velgaard OlsenOlsen JVHayward DGNilsson JKelstrup CDSchou JfalseComprehensive identification of SUMO2/3 targets and their dynamics during mitosis.During mitosis large alterations in cellular structures occur rapidly, which to a large extent is regulated by post-translational modification of proteins. Modification of proteins with the small ubiquitin-related protein SUMO2/3 regulates mitotic progression, but few mitotic targets have been identified so far. To deepen our understanding of SUMO2/3 during this window of the cell cycle, we undertook a comprehensive proteomic characterization of SUMO2/3 modified proteins in mitosis and upon mitotic exit. We developed an efficient tandem affinity purification strategy of SUMO2/3 modified proteins from mitotic cells. Combining this purification strategy with cell synchronization procedures and quantitative mass spectrometry allowed for the mapping of numerous novel targets and their dynamics2014-01-01T00:00:00Z20142025-06-01T00:33:17.585Z2025-06-01T00:33:17.585ZS-EPMC40740682497188810.1371/journal.pone.0100692