Detecting endogenous SUMO targets in mammalian cells and tissues

Nat Struct Mol Biol. 2013 Apr;20(4):525-31. doi: 10.1038/nsmb.2526. Epub 2013 Mar 17.


SUMOylation is an essential modification that regulates hundreds of proteins in eukaryotic cells. Owing to its dynamic nature and low steady-state levels, endogenous SUMOylation is challenging to detect. Here, we present a method that allows efficient enrichment and identification of endogenous targets of SUMO1 and the nearly identical SUMO2 and 3 (SUMO 2/3) from vertebrate cells and complex organ tissue. Using monoclonal antibodies for which we mapped the epitope, we enriched SUMOylated proteins by immunoprecipitation and peptide elution. We used this approach in combination with MS to identify SUMOylated proteins, which resulted in the first direct comparison of the endogenous SUMO1- and SUMO2/3-modified proteome in mammalian cells, to our knowledge. This protocol provides an affordable and feasible tool to investigate endogenous SUMOylation in primary cells, tissues and organs, and it will facilitate understanding of SUMO's role in physiology and disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / immunology
  • Cells, Cultured
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Mammals
  • Molecular Sequence Data
  • Sequence Homology, Amino Acid
  • Small Ubiquitin-Related Modifier Proteins / chemistry
  • Small Ubiquitin-Related Modifier Proteins / immunology
  • Small Ubiquitin-Related Modifier Proteins / metabolism*


  • Antibodies, Monoclonal
  • Small Ubiquitin-Related Modifier Proteins