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.
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Data availability
The backbone 1H, 13C and 15N resonance assignment of apo Ubiquitin Specific Protease 7 catalytic domain (residues 208–554) was deposited in Biological Magnetic Resonance Bank under accession code 51912. The assignment for the complex with compound 21 (O’Dowd et al. 2018a) was deposited under code 51913.
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This work was supported by Innovate UK (Knowledge Transfer Partnership 11447).
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M.J.P., M.J.C., J.P.W. and M.J.W. initiated the project and designed the experimental protocol. I.L, A.S., J.K. and K.T. expressed and purified protein. M.J.P. performed hydrogen exchange/protein re-folding. M.J.P. M.J.C and H.R.W.D. recorded spectra. W.A. performed resonance assignments, analysed the results, wrote the manuscript and prepared the figures. All authors reviewed the manuscript.
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Pandya, M.J., Augustyniak, W., Cliff, M.J. et al. 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. Biomol NMR Assign (2024). https://doi.org/10.1007/s12104-024-10165-7
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DOI: https://doi.org/10.1007/s12104-024-10165-7