Dissecting SUMO Dynamics by Mass Spectrometry.
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.
Department of Biophysics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, Warszawa, 02-106, Poland. drabikowski@ibb.waw.pl.
Krzysztof
K
Drabikowski
Department of Biophysics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, Warszawa, 02-106, Poland.
Michał
M
Dadlez
D000818:Animals; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D011499:Protein Processing, Post-Translational / Q000235:genetics / Q000502:physiology; D040901:Proteomics; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics / Q000378:metabolism*; D058207:Sumoylation / Q000235:genetics / Q000502:physiology
D025841:Small Ubiquitin-Related Modifier Proteins
Label-free proteomics; Posttranslational modifications; SUMO; Stress response; Ubiquitin like proteins