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Hokkaido University Collection of Scholarly and Academic Papers >
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Participation of the surface structure of pharaonis phoborhodopsin, ppR and its A149S and A149V mutants, consisting of the C-terminal α-helix and E-F loop, in the complex-formation with the cognate transducer pHtrII, as revealed by site-directed 13C solid-state NMR
| Title: | Participation of the surface structure of pharaonis phoborhodopsin, ppR and its A149S and A149V mutants, consisting of the C-terminal α-helix and E-F loop, in the complex-formation with the cognate transducer pHtrII, as revealed by site-directed 13C solid-state NMR |
| Authors: | Kawamura, Izuru Browse this author | | Ikeda, Yoichi Browse this author | | Sudo, Yuki Browse this author | | Iwamoto, Masayuki Browse this author | | Shimono, Kazumi Browse this author | | Yamaguchi, Satoru Browse this author | | Tuzi, Satoru Browse this author | | Saitô, Hazime Browse this author | | Kamo, Naoki Browse this author | | Naito, Akira Browse this author |
| Issue Date: | Mar-2007 |
| Publisher: | Blackwell Publishing |
| Journal Title: | Photochemistry and Photobiology |
| Volume: | 83 |
| Issue: | 2 |
| Start Page: | 339 |
| End Page: | 345 |
| Publisher DOI: | 10.1562/2006-06-20-RA-940 |
| Abstract: | We have recorded 13C solid state NMR spectra of [3-13C]Ala-labeled pharaonis phoborhodopsin ppR, and its mutants, A149S and A149V, complexed with the cognate transducer pHtrII (1-159), to gain insight into a possible role of their cytoplasmic surface structure including the C-terminal α-helix and E-F loop for stabilization of the 2:2 complex, by both cross-polarization magic angle spinning (CP-MAS) and dipolar decoupled-magic angle spinning (DD-MAS) NMR techniques. We found that 13C CP-MAS NMR spectra of [3-13C]Ala-ppR, A149S, and A149V complexed with the transducer pHtrII are very similar, reflecting their conformation and dynamics changes caused by mutual interactions through the transmembrane α-helical surfaces. In contrast, their DD-MAS NMR spectral features are quite different between [3-13C]Ala- A149S and A149V in the complexes with pHtrII: 13C DD-MAS NMR spectrum of [3-13C]Ala-A149S complex is rather similar to that of the uncomplexed form, while the corresponding spectral feature of A149V complex is similar to that of ppR complex in the C-terminal tip region. This is because more flexible surface structure detected by the DD-MAS NMR spectra are more directly influenced by the dynamics changes than the CP-MAS NMR. It turned out, therefore, that an altered surface structure of A149S resulted in destabilized complex as viewed from the 13C NMR spectrum of the surface areas, probably because of modified conformation at the corner of the helix E in addition to the change of hydropathy. It is, therefore, concluded that the surface structure of ppR including the C-terminal α-helix and the E-F loops is directly involved in the stabilization of the complex through conformational stability of the helix E. |
| Rights: | Author Posting. © The Authors 2007 This is the author's version of the work. It is posted here for personal use, not for redistribution. The definitive version was published in Photochemistry and Photobiology, Volume 83, Issue 2, Page 339-345. http://dx.doi.org/10.1562/2006-06-20-RA-94 |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/33765 |
| 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: 加茂 直樹
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