Analysis of SUMOylated proteins using SUMO-traps

Elisa da Silva-Ferrada (1, 2) , Wendy Xolalpa (1, 2) , Valérie Lang (2) , Fabienne Aillet (2) , Itziar Martin-Ruiz (1) , Carlos F de La Cruz-Herrera (3) , Fernando Lopitz-Otsoa (1) , Arkaitz Carracedo (1, 4, 5) , Seth J Goldenberg , Carmen Rivas (3) , Patrick England (6, 7) , Manuel S Rodríguez (1, 2)

1 CIC bioGUNE - Center for Cooperative Research in Biosciences = Centro de Investigación cooperativa en biociencia (Derio, Biscay, Espagne)
2 Inbiomed [Spain]
3 CSIC - Consejo Superior de Investigaciones Cientificas [España] = Spanish National Research Council [Spain]
4 Ikerbasque - Basque Foundation for Science
5 UPV / EHU - University of the Basque Country = Euskal Herriko Unibertsitatea
6 IP - Institut Pasteur [Paris]
7 Biophysique des macromolécules et leurs interactions

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.

Domaines: Sciences du Vivant [q-bio]

Elisa da Silva-Ferrada, Wendy Xolalpa, Valérie Lang, Fabienne Aillet, Itziar Martin-Ruiz, et al.. Analysis of SUMOylated proteins using SUMO-traps. Scientific Reports, 2013, 3 (1), pp.1690. ⟨10.1038/srep01690⟩. ⟨pasteur-03525526⟩