Detection and Analysis of SUMOylation Substrates In Vitro and In Vivo.
SUMOylation is a widely used protein posttranslational mechanism capable of regulating substrates localization, stability, and/or activity. Identification and characterization of bona fide SUMO substrates is a laborious task but its discovery can shed light to exquisite and crucial regulatory signaling events occurring within the cell. Experiments performed in the SUMOylation field often demand a good understanding of the putative substrate's function and necessitate a solid knowledge regarding both in vitro and in vivo approaches. This contribution offers a simplified view into some of the most common experiments performed in biochemical and cell biological research of the SUMO pathway in mammalian systems. It also summarizes and updates well established protocols and tricks in order to improve the likelihood to obtain reliable and reproducible results.
Department of Structural Biology, Vlaams Instituut voor Biotechnologie (VIB), Vrije Universiteit Brussel (VUB), Pleinlaan 2, Brussels, 1050, Belgium.
Cesyen
C
Cedeño
Laboratory of Pathophysiological Cell Signaling (PACS), Department of Biology, Faculty of Sciences and Bioengineering Sciences, Vrije Universiteit Brussel (VUB), Pleinlaan 2, Brussels, 1050, Belgium.
Esther
E
La Monaca
Laboratory of Pathophysiological Cell Signaling (PACS), Department of Biology, Faculty of Sciences and Bioengineering Sciences, Vrije Universiteit Brussel (VUB), Pleinlaan 2, Brussels, 1050, Belgium.
Mara
M
Esposito
Laboratory of Pathophysiological Cell Signaling (PACS), Department of Biology, Faculty of Sciences and Bioengineering Sciences, Vrije Universiteit Brussel (VUB), Pleinlaan 2, Brussels, 1050, Belgium. ggutierr@vub.ac.be.
Gustavo J
GJ
Gutierrez
D000818:Animals; D006801:Humans; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics / Q000378:metabolism*; D058207:Sumoylation / Q000235:genetics / Q000502:physiology*
D025841:Small Ubiquitin-Related Modifier Proteins
SUMO pathway; SUMO substrates; SUMOylation