Despite significant advancements in our understanding of ubiquitin-mediated signaling, the influence of the intracellular environment on the formation of transient ubiquitin-partner complexes remains poorly explored. In our work, we introduce macromolecular crowding as a first level of complexity toward the imitation of a cellular environment in the study of such interactions. Using NMR spectroscopy, we find that the stereospecific complex of ubiquitin and the ubiquitin-associated domain (UBA) is minimally perturbed by the crowding agent Ficoll. However, in addition to the primary canonical recognition patch on ubiquitin, secondary patches are identified, indicating that in cell-mimicking crowded solution, UBA contacts ubiquitin at multiple sites.

Identification of primary and secondary UBA footprints on the surface of ubiquitin in cell-mimicking crowded solution

MUNARI, FRANCESCA;Bortot, Andrea;ZANZONI, Serena;D'ONOFRIO, Mariapina;ASSFALG, Michael
2017

Abstract

Despite significant advancements in our understanding of ubiquitin-mediated signaling, the influence of the intracellular environment on the formation of transient ubiquitin-partner complexes remains poorly explored. In our work, we introduce macromolecular crowding as a first level of complexity toward the imitation of a cellular environment in the study of such interactions. Using NMR spectroscopy, we find that the stereospecific complex of ubiquitin and the ubiquitin-associated domain (UBA) is minimally perturbed by the crowding agent Ficoll. However, in addition to the primary canonical recognition patch on ubiquitin, secondary patches are identified, indicating that in cell-mimicking crowded solution, UBA contacts ubiquitin at multiple sites.
NMR spectroscopy; biomolecular recognition; macromolecular crowding; paramagnetic relaxation enhancement; ubiquitin; ubiquitin-associated domain
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/960568
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