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Uncovering the SUMOylation and ubiquitylation crosstalk in human cells using sequential peptide immunopurification.
- Source :
-
Nature communications [Nat Commun] 2017 Jan 18; Vol. 8, pp. 14109. Date of Electronic Publication: 2017 Jan 18. - Publication Year :
- 2017
-
Abstract
- Crosstalk between the SUMO and ubiquitin pathways has recently been reported. However, no approach currently exists to determine the interrelationship between these modifications. Here, we report an optimized immunoaffinity method that permits the study of both protein ubiquitylation and SUMOylation from a single sample. This method enables the unprecedented identification of 10,388 SUMO sites in HEK293 cells. The sequential use of SUMO and ubiquitin remnant immunoaffinity purification facilitates the dynamic profiling of SUMOylated and ubiquitylated proteins in HEK293 cells treated with the proteasome inhibitor MG132. Quantitative proteomic analyses reveals crosstalk between substrates that control protein degradation, and highlights co-regulation of SUMOylation and ubiquitylation levels on deubiquitinase enzymes and the SUMOylation of proteasome subunits. The SUMOylation of the proteasome affects its recruitment to promyelocytic leukemia protein (PML) nuclear bodies, and PML lacking the SUMO interacting motif fails to colocalize with SUMOylated proteasome further demonstrating that this motif is required for PML catabolism.
- Subjects :
- Amino Acid Motifs
HEK293 Cells
Humans
Promyelocytic Leukemia Protein chemistry
Promyelocytic Leukemia Protein genetics
Promyelocytic Leukemia Protein metabolism
Proteasome Endopeptidase Complex genetics
Proteasome Endopeptidase Complex metabolism
Protein Interaction Maps
Proteins chemistry
Proteins genetics
Proteolysis
Sumoylation
Ubiquitin chemistry
Ubiquitination
Chromatography, Affinity methods
Peptides chemistry
Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2041-1723
- Volume :
- 8
- Database :
- MEDLINE
- Journal :
- Nature communications
- Publication Type :
- Academic Journal
- Accession number :
- 28098164
- Full Text :
- https://doi.org/10.1038/ncomms14109