title: Real-Time Surface Plasmon Resonance (SPR) for the Analysis of Interactions Between SUMO Traps and Mono- or PolySUMO Moieties.
abstract: Isolating endogenous SUMOylated proteins is a challenging task due to the high reversibility of this posttranslational modification. We have shown that SUMO traps are useful tools for the enrichment and isolation of proteins modified by SUMO in vitro and in vivo. To characterize the affinity and specificity of different SUMO chains for these traps, that are based on SUMO-interacting motifs, we have used real-time surface plasmon resonance (SPR), which allows a label-free analysis of protein/protein interactions. Here, a protocol to determine the affinities of multivalent SUMO traps for polySUMO chains or mono-SUMO molecules by SPR is presented.
authors.affiliation: Centro de Investigación sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pública, 62100, Cuernavaca, Morelos, Mexico.
authors.forename: Wendy
authors.initials: W
authors.lastname: Xolalpa
authors.affiliation: INBIOMED, San Sebastian, Spain.
authors.forename: Manuel S
authors.initials: MS
authors.lastname: Rodriguez
authors.affiliation: Plate-forme de Biophysique des Macromolécules et de leurs Interactions, Institut Pasteur, 75724, Paris, France. Patrick.england@pasteur.fr.|CNRS-UMR 3528, 75724, Paris, France. Patrick.england@pasteur.fr.
authors.forename: Patrick
authors.initials: P
authors.lastname: England
mesh_terms: D000906:Antibodies / Q000737:chemistry; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D046208:Microchip Analytical Procedures; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D025842:SUMO-1 Protein / Q000235:genetics / Q000378:metabolism*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics / Q000378:metabolism*; D058207:Sumoylation; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D000906:Antibodies; D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein; C495501:SUMO1 protein, human; C117909:SUMO2 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; D005982:Glutathione Transferase
keywords: Affinity constants; Avidity; Multivalence; Protein interactions; SPR; SUMO-binding entities