Proteostasis pp 291-298 | Cite as

Dissecting SUMO Dynamics by Mass Spectrometry

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1449)

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.

Key words

SUMO Ubiquitin like proteins Posttranslational modifications Label-free proteomics Stress response 

Notes

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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of BiophysicsInstitute of Biochemistry and Biophysics, Polish Academy of SciencesWarszawaPoland

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