Abstract
Two new 3D HN-based experiments are proposed for backbone assignment of large disordered proteins. The spectra obtained with the new pulse schemes are free of redundant diagonal peaks (HiNi–Ni) and provide sequential correlations (HiNi–Ni+1 and HiNi–Ni−1) not only between adjacent non-proline residues but also between non-proline and proline residues. The experiments have been demonstrated on an intrinsically disordered protein with 306 amino acids including 64 proline residues. Using the two experiments, we obtained nearly complete assignments of backbone amides and proline 15N spins except for 4 proline and 4 non-proline residues.
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Acknowledgments
The authors thank Prof. Sawada in Mechanobiology Institute, National University of Singapore for providing the p130CasSD construct. This work is supported by a grant from Singapore Ministry of Education Academic Research Fund Tier 1 (R154000555112).
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Liu, X., Yang, D. HN(CA)N and HN(COCA)N experiments for assignment of large disordered proteins. J Biomol NMR 57, 83–89 (2013). https://doi.org/10.1007/s10858-013-9783-1
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DOI: https://doi.org/10.1007/s10858-013-9783-1