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Journal Article

Detecting endogenous SUMO targets in mammalian cells and tissues.

MPS-Authors
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Karaca,  S.
Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society;

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Hsiao,  H. H.
Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons15947

Urlaub,  H.
Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society;

Fulltext (public)

1740152.pdf
(Publisher version), 903KB

Supplementary Material (public)

1740152_Supplement_1.pdf
(Supplementary material), 3MB

Citation

Becker, J., Barysch, S. V., Karaca, S., Dittmer, C., Hsiao, H. H., Diaz, M. B., et al. (2013). Detecting endogenous SUMO targets in mammalian cells and tissues. Nature Structural and Molecular Biology, 20(4), 525-531. doi:10.1038/nsmb.2526.


Cite as: http://hdl.handle.net/11858/00-001M-0000-000E-FBF3-6
Abstract
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.