Real-Time Surface Plasmon Resonance (SPR) for the Analysis of Interactions Between SUMO Traps and Mono- or PolySUMO Moieties

Wendy Xolalpa (1) , Manuel S Rodriguez (2) , Patrick England (3)

1 Centro de Investigacion Sobre Enfermedades Infecciosas
2 Inbiomed [Spain]
3 Biophysique des macromolécules et leurs interactions

Isolating endogenous SUMOylated proteins is a challenging task due to the high reversibility of this posttranslational modifi cation. We have shown that SUMO traps are useful tools for the enrichment and isolation of proteins modifi ed by SUMO in vitro and in vivo. To characterize the affi nity and specifi city of different SUMO chains for these traps, that are based on SUMO-interacting motifs, we have used realtime surface plasmon resonance (SPR), which allows a label-free analysis of protein/protein interactions. Here, a protocol to determine the affi nities of multivalent SUMO traps for polySUMO chains or mono-SUMO molecules by SPR is presented.

Mots clés: SPR; Protein interactions; SUMO-binding entities; Affi nity constants; Avidity; Multivalence; Affinity constants

Domaines: Sciences du Vivant [q-bio]

Wendy Xolalpa, Manuel S Rodriguez, Patrick England. Real-Time Surface Plasmon Resonance (SPR) for the Analysis of Interactions Between SUMO Traps and Mono- or PolySUMO Moieties. SUMO, 1475, Springer New York; Springer New York, pp.99 - 107, 2016, Methods in Molecular Biology, ⟨10.1007/978-1-4939-6358-4_7⟩. ⟨pasteur-03525436⟩