Abstract
YibK is a tRNA methyltransferase from Haemophilus influenzae, which forms a stable homodimer in solution and contains a deep trefoil 31 knot encompassing the C-terminal helix that threads through a long loop. It has been a model system for investigating knotted protein folding pathways. Recent data have shown that the polypeptide chain of YibK remains loosely knotted under highly denaturing conditions. Here, we report 1H, 13C and 15N chemical shift assignments for YibK and its variant in the presence of 8 M urea. This work forms the basis for further analysis using NMR techniques such as paramagnetic relaxation enhancement, residual dipolar couplings and spin-relaxation dynamics analysis.
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Acknowledgments
The NMR spectra were obtained at the Core Facility for Protein Structural Analysis supported by National Core Facility Program for Biotechnology. STDH is a recipient of the Career Development Award (CDA—00025/2010-C) from the International Human Frontier Science Program and is supported by funding from the National Science Council (100-2113-M-001-031-MY2 and 101-2627-M-001-004) and Academia Sinica, Taiwan.
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Hsieh, SJ.M., Mallam, A.L., Jackson, S.E. et al. Backbone NMR assignments of a topologically knotted protein in urea-denatured state. Biomol NMR Assign 8, 439–442 (2014). https://doi.org/10.1007/s12104-013-9510-6
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DOI: https://doi.org/10.1007/s12104-013-9510-6