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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Life Science / Faculty of Advanced Life Science >
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Convenient method for resolving degeneracies due to symmetry of the magnetic susceptibility tensor and its application to pseudo contact shift-based protein-protein complex structure determination
| Title: | Convenient method for resolving degeneracies due to symmetry of the magnetic susceptibility tensor and its application to pseudo contact shift-based protein-protein complex structure determination |
| Authors: | Kobashigawa, Yoshihiro Browse this author | | Saio, Tomohide Browse this author | | Ushio, Masahiro Browse this author | | Sekiguchi, Mitsuhiro Browse this author | | Yokochi, Masashi Browse this author | | Ogura, Kenji Browse this author →KAKEN DB | | Inagaki, Fuyuhiko Browse this author →KAKEN DB |
| Keywords: | Differential scanning fluorometry | | Pseudo contact shift | | Lanthanide binding tag | | FKBP12 | | FRB | | mTOR |
| Issue Date: | May-2012 |
| Publisher: | Springer Netherlands |
| Journal Title: | Journal of Biomolecular NMR |
| Volume: | 53 |
| Issue: | 1 |
| Start Page: | 53 |
| End Page: | 63 |
| Publisher DOI: | 10.1007/s10858-012-9623-8 |
| Abstract: | Pseudo contact shifts (PCSs) induced by paramagnetic lanthanide ions fixed in a protein frame provide long-range distance and angular information, and are valuable for the structure determination of protein-protein and protein-ligand complexes. We have been developing a lanthanide-binding peptide tag (hereafter LBT) anchored at two points via a peptide bond and a disulfide bond to the target proteins. However, the magnetic susceptibility tensor displays symmetry, which can cause multiple degenerated solutions in a structure calculation based solely on PCSs. Here we show a convenient method for resolving this degeneracy by changing the spacer length between the LBT and target protein. We applied this approach to PCS-based rigid body docking between the FKBP12-rapamycin complex and the mTOR FRB domain, and demonstrated that degeneracy could be resolved using the PCS restraints obtained from two-point anchored LBT with two different spacer lengths. The present strategy will markedly increase the usefulness of two-point anchored LBT for protein complex structure determination. |
| Type: | article |
| URI: | http://hdl.handle.net/2115/49373 |
| 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: 稲垣 冬彦
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