| 1.Research Institution | Kyoto University | |
| 2.Research Area | Life Sciences | |
| 3.Research Field | Structure and Functional Control mechanism of Biomolecules | |
| 4.Term of Project | FY1996〜FY2000 | |
| 5.Project Number | 96L00502 | |
| 6.Title of Project | Structural Study of Molecular Interaction in Living Organism |
| Name | Institution,Department | Title of Position |
| Yoshinori Fujiyoshi | Graduate School of Science, Kyoto University | Professor |
8.Core Members
| Names | Institution,Department | Title of Position |
| Hiromichi Sato | Graduate School of Pharmaceutical Science, Kyoto University | Professor |
9.Cooperating Researchers
| Names | Institution,Department | Title of Position |
| Kaoru Mitsuoka | Graduate School of Science, Kyoto University | Assistant |
10.Summary of Research Results
|
The project was executed by focusing on structural study of membrane proteins including channels through technical and methodological development for structure analysis of membrane proteins which were involved in important biological functions such as signal transduction. The following are the main results in this project. 1) A structure of water channel, aquaporin-1 (AQP-1) was analysed at a resolution of 6Å by electron crystallography (Nature, 387, 624-627 (1997).). It was suggested that charge shifts on bacteriorhodopsin molecule were discriminated based on the structure analysis at 3Å resolution by electron crystallography (Nature, 389, 206-211 (1997). & J. Mol. Biol., 286, 861-882 (1999).). An atomic structure of AQP-1 was analysed and the model provides a possible molecular explanation to longstanding puzzles in physiology; i) How does mercurial ion inhibit the water permeability through the channel? ii) How does the channel achieve marked selectivity? iii) How does the channel achieve fast water transport? iv) How does the molecule block the proton transport despite the high-speed water flow? (Nature, 405, 599-605 (2000).). 2) The structure of Nicotinic acetylcholine receptor, which was found in the postsynaptic membranes of muscle cells, was analysed at 4.6Å resolution (J. Mol. Biol., 288, 765-786 (1999).). 3) Three-dimensional structure of voltage sensitive Na+-channel, which was purified from electric eel, was analysed at 19Å resolution based on single particle analysis using an electron cryo-microscope (Nature, 409, 1047-1051 (2001).). 4) An electron microscope with helium cooled stage was improved by modifying the microscopic system including the stage which was developed by Fujiyoshi et al. in 1986. 5) Functional studies of G-protein coupled receptors were proceeded through developing a system for over-expression and purification (for example, Biochemistry, 39, 686-692 (2000).). |
11.Key Words
(1)membrane proteins、(2)three-dimensional structure、(3)water channel
(4)Nicotinic acetylcholine receptor、(5)bacteriorhodopsin、(6)voltage sensitive Na+-channel
(7)G-protein coupled receptors、(8)electron crystallography、(9)electron cryo-microscope
12.References
| Author | Title of Article | |||
| A. H. Iwane, I. Ikeda, Y. Kimura, Y. Fujiyoshi, K. Aitendorf and W. Epstein | Two-Dimensional Crystal of the Kdp-ATPase of Escherichia coli. | |||
| Journal | Volume | Year | Pages Concerned | |
| FEBS Letters | 396 | 1996 | 172-176 | |
| Author | Title of Article | |||
| Y. M. Kiyosue, F. Vonderviszt, I. Yamashita, Y. Fujiyoshi and K. Namba. | Direct Interaction of Flagellin Termini Essential for polymorphic Ability of Flagellar Filament. | |||
| Journal | Volume | Year | Pages Concerned | |
| Proc. Natl. Acad. Sci. USA | 93 | 1996 | 15108-15113 | |
| Author | Title of Article | |||
| T. Walz, T. Hirai, K. Murata, B. L. Smith, J. B. Heymann, K. Mitsuoka, Y. Fujiyoshi, P. Agre and A. Engel | Theree-Dimensional Structure of Aquaporin 1. | |||
| Journal | Volume | Year | Pages Concerned | |
| Nature | 387 | 1997 | 624-627 | |
| Author | Title of Article | |||
| Y. Kimura, D. G. Vassylyev, A. Miyazawa, A. Kidera, M. Matsushima, K. Mitsuoka, K. Murata, T. Hirai and Y. Fujiyoshi | Surface of bacteriorhodopsin revealed by high resolution electron crystallography. | |||
| Journal | Volume | Year | Pages Concerned | |
| Nature | 389 | 1997 | 206-211 | |
| Author | Title of Article | |||
| H. Hebert, I. Schmidt-Krey, R. Morgenstem, K. Murata, T. Hirai, K. Mitsuoka and Y. Fujiyoshi | The 3.0Å projection structure of microsomal glutathione transferase as determind by electron crystallography of p21212 two-dimenstional crystals. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Mol. Biol. | 271 | 1997 | 751-758 | |
| Author | Title of Article | |||
| Y. Kimura, D. G. Vassylyev, A. Miyazawa, A. Kidera, M. Matsushima, K. Mitsuoka, K. Murata, T. Hirai and Y. Fujiyoshi | High resolution structure of bacteriorhodopsin determined by electron crystallography. | |||
| Journal | Volume | Year | Pages Concerned | |
| Photochemistry and Photobiology | 66 | 1997 | 764-767 | |
| Author | Title of Article | |||
| T. Doi, Y. Hioaki, I. Arimoto, Y. Fujiyoshi, T. Okamoto, M. Satoh and Y. Furichi | Characterization of human endothelin ETB receptor and mutants expressed in insect cells. | |||
| Journal | Volume | Year | Pages Concerned | |
| Eur. J. Biochem. | 248 | 1997 | 139-148 | |
| Author | Title of Article | |||
| Y. Zhu, A. M. Hemmings, K. Iwasaki, Y. Fujiyoshi, B. Zhong, J. Yan, M. Isogai and T. Omura | Details of the arrangement of the outer capsid of rice dwarf phytoreovirue as visualized by two-dimensional crystallograohy. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Virol. | 71 | 1997 | 8899-8901 | |
| Author | Title of Article | |||
| K. Mitsuoka, K. Murata, Y. Kimura, K. Namba and Y. Fujiyoshi | Examination of the leafscan 45, a line-illuminating micro-densitometer for its use in electron crystallography. | |||
| Journal | Volume | Year | Pages Concerned | |
| Uitramicroscopy | 68 | 1997 | 109-121 | |
| Author | Title of Article | |||
| T. Ohtaki, K. Ogi, Y. Masudall, K. Mitsuoka, Y. Fujiyoshi, C. Kitada, H. Sawada, H. Oda and M. Fujino. | Expression, purification and reconstitution of receptor for pituitary adenylate cyclase-activating polypeptide. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Bipl. Chem. | 273 | 1998 | 15464-15473 | |
| Author | Title of Article | |||
| Y. M. Kiyosue, I. Yamashita, Y. Fujiyoshi, S. Yamaguchi and K. Namba | Role of the outermost subdomain of salmonella flagelline in the filament structure revealed by electron cryomicroscopy. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Mol. Biol. | 284 | 1998 | 521-530 | |
| Author | Title of Article | |||
| Y. Fujiyoshi | The structural study of membrance proteins by electron crystallography. | |||
| Journal | Volume | Year | Pages Concerned | |
| Adv. Biophys. | 35 | 1998 | 25-80 | |
| Author | Title of Article | |||
| I. Ikeda, T. Odahara, K. Mitsuoka, Y. Fujiyoshi and K. Murata | Projection map of the reaction center-light harvesting complex from rhodopseudomanas viridis at 10 A resolution. | |||
| Journal | Volume | Year | Pages Concerned | |
| FEBS Letters. | 425 | 1998 | 505-508 | |
| Author | Title of Article | |||
| K. Mitsuoka, T. Hirai, K. Murata, A. Miyazawa, A. Kidera, Y. Kimura and Y. Fujiyoshi | The structure of bacteriorhodopsin at 3.0 Å rsolution based on electron crystallography: Implication of the charge distribution. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Mol. Biol. | 286 | 1999 | 861-882 | |
| Author | Title of Article | |||
| Y. Hiroaki, K. Mitsuoka, Y. Cheng H. Hiroaki and Y. Fujiyoshi | Influence of various nucleotides on the in situ crystallization of Ca2+ -ATPase. | |||
| Journal | Volume | Year | Pages Concerned | |
| Biochemica et Biophysica Acta | 1415 | 1999 | 361-368 | |
| Author | Title of Article | |||
| T. Doi, H. Sugimoto, I. Arimoto, Y. Hiroaki and Y. Fujiyoshi | Interactions of endothelin receptor subtypes A and B with Gi, Go and Gq in reconstituted phospholipid vesicles. | |||
| Journal | Volume | Year | Pages Concerned | |
| Biochemistry | 38 | 1999 | 3090-3099 | |
| Author | Title of Article | |||
| Y. Fujiyoshi | Electron crystallography of membrance proteins. | |||
| Journal | Volume | Year | Pages Concerned | |
| G. protein-coupled receptors(CRC Press: Edited by T. Haga and G. Berstein) | 1999 | 323-354 | ||
| Author | Title of Article | |||
| A. Miyazawa, Y. Fujiyoshi, M. Stowell and N. Unwin | Nicotinic Acetylcholine Receptor at 4.6Å Resolution: Transverse Tunnels in the Channel Wall. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Mol. Biol. | 288 | 1999 | 765-786 | |
| Author | Title of Article | |||
| T. Hirai, K. Murata, K. Mitsuoka, Y. Kimura and Y. Fujiyoshi | Trehalose embedding technique for high-resolution electron crystallography. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Electron Microsc. | 48 | 1999 | 653-658 | |
| Author | Title of Article | |||
| Y. Fujiyoshi | Molecular structure of proton pump revealed with electron crystallography. | |||
| Journal | Volume | Year | Pages Concerned | |
| The FASEB Journal. | 1999 | S191-s194 | ||
| Author | Title of Article | |||
| K. Mitsuoka, K. Murata, T. Walz, T. Hirai, P. Agre, J. B. Heymann, A. Engel and Y. Fujiyoshi. | The Structure of Aquaporin-1 4.5Å Resolution Reveals Short α-Helices in the Center of the Monomer. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Structure Biol. | 128 | 1999 | 34-43 | |
| Author | Title of Article | |||
| F. Imamura, I. Arimoto, Y. Fujiyoshi and T. Doi | W276 Mutation in the Endothelin Receptor subtype B Imaparis Gq Coupling but not Gi or Go Coupling. | |||
| Journal | Volume | Year | Pages Concerned | |
| Biochemistry. | 39 | 2000 | 686-692 | |
| Author | Title of Article | |||
| Y. Tahara, A. Oshima, T. Hirai, K. Mitsuoka, Y. fujiyoshi and Y. Hayashi. | The 11Å resolution projection map of Na+/K+-ATPase calculated by application pf single particle anaysis to two-dimensional crystal images. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Electron Microsc. | 49 | 2000 | 583-587 | |
| Author | Title of Article | |||
| B. L. de Groot, J. B. Heymann, A. Engel, K. Mitsuoka, Y. Fujioshi and H. Grubmueller | The fold of human aquaporinl | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Mol. Biol. | 300 | 2000 | 987-994 | |
| Author | Title of Article | |||
| K. Murata, K. Mitsuoka, T. Hirai, T. Walz, P. Agre, J. B. Heymann, A. Engel and Y. Fujiyoshi | Structure determinants of water permeation through aqaporin-1. | |||
| Journal | Volume | Year | Pages Concerned | |
| Nature. | 407 | 2000 | 599-605 | |
| Author | Title of Article | |||
| I. Schmidt-Krey, K. Mitsuoka, T. Hirai, K. Murata, Y. Cheng, Y. Fujiyoshi, R. Morgenstern and H. Hebert | The three-dimensional map of microsomal glutathione transferase 1 at 6 Å resolution | |||
| Journal | Volume | Year | Pages Concerned | |
| EMBO Journal. | 19 | 2000 | 6311-6316 | |
| Author | Title of Article | |||
| C. Sato, Y. Ueno, K. Asai, K. Takahashi, M. Sato, A. Engel and Y. Fujiyoshi | The voltage-sensitive sodium channel is a bell-shaped molecule with several cavities. | |||
| Journal | Volume | Year | Pages Concerned | |
| Neture. | 409 | 2001 | 1047-1051 | |
| Author | Title of Article | |||
| A. Engel, Y. Fujiyoshi and P. Age | The Importance of Aquaporin Water Channel Protein structures | |||
| Journal | Volume | Year | Pages Concerned | |
| EMBO J | 19 | 2000 | 800-806 | |