Protein modification by SUMO proteins is one of the key posttranslational modifications in eukaryotes. Here, we describe a workflow to analyze SUMO dynamics in response to different stimuli, purify SUMO conjugates, and analyze the changes in SUMOylation level in organisms, tissues, or cell culture. We present a protocol for lysis in denaturing conditions that is compatible with downstream IMAC and antibody affinity purification, followed by mass spectrometry and data analysis.
SUMO Ubiquitin like proteins Posttranslational modifications Label-free proteomics Stress response
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The work was supported by National Science Centre (Narodowe Centrum Nauki) grant DEC 1/Z/2011/01/M/NZ2/02997. The authors would like to acknowledge networking support by the Proteostasis COST Action (BM1307).
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