Abstract
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.
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Acknowledgments
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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Drabikowski, K., Dadlez, M. (2016). Dissecting SUMO Dynamics by Mass Spectrometry. In: Matthiesen, R. (eds) Proteostasis. Methods in Molecular Biology, vol 1449. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3756-1_18
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DOI: https://doi.org/10.1007/978-1-4939-3756-1_18
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3754-7
Online ISBN: 978-1-4939-3756-1
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