title: Analysis of SUMOylated proteins using SUMO-traps.
abstract: SUMO-modified proteins are recognized by SUMO interacting motifs (SIMs), thus triggering diverse cellular responses. Here SIMs were used to develop SUMO-traps to capture endogenous SUMOylated proteins. Our results show that these small peptides are transferable motifs that maintain their SUMO binding capacity when fused to the heterologous carrier protein GST. The tandem disposition of SIMs increases the binding capacity of SUMO-traps to specifically interact with polySUMO but not poly-Ubiquitin chains. We demonstrate that this SUMO capturing system purifies SUMOylated proteins such as IκBα, PTEN, PML or p53 in vitro and in vivo. These properties can be used to explore the many critical functions regulated by protein SUMOylation.
authors.affiliation: Proteomics Unit, CIC bioGUNE, CIBERehd, Bd. 801A, Bizkaia Technology Park, Derio, 48160, Bizkaia, Spain.
authors.forename: Elisa
authors.initials: E
authors.lastname: Da Silva-Ferrada
authors.forename: Wendy
authors.initials: W
authors.lastname: Xolalpa
authors.forename: Valérie
authors.initials: V
authors.lastname: Lang
authors.forename: Fabienne
authors.initials: F
authors.lastname: Aillet
authors.forename: Itziar
authors.initials: I
authors.lastname: Martin-Ruiz
authors.forename: Carlos F
authors.initials: CF
authors.lastname: de la Cruz-Herrera
authors.forename: Fernando
authors.initials: F
authors.lastname: Lopitz-Otsoa
authors.forename: Arkaitz
authors.initials: A
authors.lastname: Carracedo
authors.forename: Seth J
authors.initials: SJ
authors.lastname: Goldenberg
authors.forename: Carmen
authors.initials: C
authors.lastname: Rivas
authors.forename: Patrick
authors.initials: P
authors.lastname: England
authors.forename: Manuel S
authors.initials: MS
authors.lastname: Rodríguez
mesh_terms: D006801:Humans; D025841:Small Ubiquitin-Related Modifier Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D058207:Sumoylation
chemical_list: D025841:Small Ubiquitin-Related Modifier Proteins