Journal
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Biochimie
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SUMO-Binding Entities (SUBEs) as Tools for the Enrichment, Isolation, Identification, and Characterization of the SUMO Proteome in Liver Cancer
Journal JoVE
Biochimie
This content is Free Access.
Journal JoVE Biochimie
SUMO-Binding Entities (SUBEs) as Tools for the Enrichment, Isolation, Identification, and Characterization of the SUMO Proteome in Liver Cancer

SUMO-Binding Entities (SUBEs) as Tools for the Enrichment, Isolation, Identification, and Characterization of the SUMO Proteome in Liver Cancer

DOI:
10.3791/60098-v

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08:29 min

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November 01, 2019

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, , , , , ,
1Liver Disease and Liver Metabolism Lab,CIC bioGUNE, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), 2Proteomics Platforms,CIC bioGUNE, ProteoRED-ISCIII, CIBERehd, 3Advanced Technology Institute in Life Sciences (ITAV)-CNRS-IPBS,UbiCARE

Chapitres

  • 00:04Titre
  • 01:58Cell Preparation and Lysis
  • 03:27Tissue Preparation and Lysis
  • 03:54Binding of GST-SUBEs or GST Control to Glutathione-Agarose Beads
  • 04:52GST Pull Down Assay
  • 05:59Identification and Characterization of SUMO Targets
  • 06:36Results: Western Blot and Mass Spectrometry Analysis
  • 07:33Conclusion

Summary

Traduction automatique

Traduction automatique

Here, we present a protocol to enrich, isolate, identify, and characterize proteins modified by SUMO in vivo both from human hepatoma cells and liver tumors obtained from mouse models of hepatocellular carcinoma by using SUMO-binding entities (SUBEs).

Tags

Keywords: SUMO-binding EntitiesSUBEsSUMO ProteomeLiver CancerSUMOylationProtein IsolationProtein CharacterizationProteaseTandem RepeatsSIMsTherapeutic ApproachHuman Hepatoma CellsCell LysisTrypsinization

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