Backbone 1H, 13C and 15N resonance assignment of the ubiquitin specific protease 7 catalytic domain (residues 208-554) in complex with a small molecule ligand

Abstract

The backbone ¹H, ¹³C and ¹⁵N resonance assignment of Ubiquitin Specific Protease 7 catalytic domain (residues 208-554) was performed in its complex with a small molecule ligand and in its apo form as a reference. The amide ¹H-¹⁵N signal intensities were boosted by an amide hydrogen exchange protocol, where expressed ²H, ¹³C, ¹⁵N-labeled protein was unfolded and re-folded to ensure exchange of amide deuterons to protons. The resonance assignments were used to determine chemical shift perturbations on ligand binding, which are consistent with the binding site observed by crystallography.

Keywords: Amide hydrogen exchange; Chemical shift perturbation; Ligand binding; NMR resonance assignment; USP7.