HUSCAP logo Hokkaido Univ. logo

Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Life Science / Faculty of Advanced Life Science >
Peer-reviewed Journal Articles, etc >

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

Files in This Item:
e.Proofing.pdf752.15 kBPDFView/Open
Please use this identifier to cite or link to this item:http://hdl.handle.net/2115/63937

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)

Submitter: 出村 誠

Export metadata:

OAI-PMH ( junii2 , jpcoar )

MathJax is now OFF:


 

 - Hokkaido University