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Structure determination of uniformly C-13, N-15 labeled protein using qualitative distance restraints from MAS solid-state C-13-NMR observed paramagnetic relaxation enhancement
| Title: | Structure determination of uniformly C-13, N-15 labeled protein using qualitative distance restraints from MAS solid-state C-13-NMR observed paramagnetic relaxation enhancement |
| Authors: | Tamaki, Hajime Browse this author | | Egawa, Ayako Browse this author | | Kido, Kouki Browse this author | | Kameda, Tomoshi Browse this author | | Kamiya, Masakatsu Browse this author | | Kikukawa, Takashi Browse this author | | Aizawa, Tomoyasu Browse this author | | Fujiwara, Toshimichi Browse this author | | Demura, Makoto Browse this author |
| Keywords: | Solid-state NMR | | Magic-angle spinning | | Protein structure | | Paramagnetic relaxation enhancement | | CS-Rosetta |
| Issue Date: | Jan-2016 |
| Publisher: | Springer |
| Journal Title: | Journal of biomolecular NMR |
| Volume: | 64 |
| Issue: | 1 |
| Start Page: | 87 |
| End Page: | 101 |
| Publisher DOI: | 10.1007/s10858-015-0010-0 |
| PMID: | 26728076 |
| Abstract: | Magic angle spinning (MAS) solid-state nuclear magnetic resonance (NMR) is a powerful method for structure determination of insoluble biomolecules. However, structure determination by MAS solid-state NMR remains challenging because it is difficult to obtain a sufficient amount of distance restraints owing to spectral complexity. Collection of distance restraints from paramagnetic relaxation enhancement (PRE) is a promising approach to alleviate this barrier. However, the precision of distance restraints provided by PRE is limited in solid-state NMR because of incomplete averaged interactions and intermolecular PREs. In this report, the backbone structure of the B1 domain of streptococcal protein G (GB1) has been successfully determined by combining the CS-Rosetta protocol and qualitative PRE restraints. The derived structure has a C alpha RMSD of 1.49 angstrom relative to the X-ray structure. It is noteworthy that our protocol can determine the correct structure from only three cysteine-EDTA-Mn2+ mutants because this number of PRE sites is insufficient when using a conventional structure calculation method based on restrained molecular dynamics and simulated annealing. This study shows that qualitative PRE restraints can be employed effectively for protein structure determination from a limited conformational sampling space using a protein fragment library. |
| Rights: | The final publication is available at www.springerlink.com via http://dx.doi.org/10.1007/s10858-015-0010-0. |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/63937 |
| Appears in Collections: | 生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 出村 誠
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