2009 Fiscal Year Final Research Report
Structural analysis of SecA-SecYE complex from Thermus thermophilus
| Project/Area Number |
19370085
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Cell biology
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| Research Institution | Kyoto University |
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Principal Investigator |
MORI Hiroyuki Kyoto University, ウイルス研究所, 准教授 (10243271)
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| Project Period (FY) |
2007 – 2009
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| Keywords | 生体膜 |
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| Research Abstract |
Evolutionarily conserved membrane protein complex, the Sec translocon, functions as a protein-conducting channel to facilitate secretory protein export and membrane protein integration. In this project, we determined the first atomic resolution crystal structure of the SecYE translocon from a SecA (bacterial translocation motor)-containing organism, Thermus thermophilus, in complex with an anti-SecY Fab fragment. The structure has revealed a significant conformational change from the previously determined archaeal SecYEβ structure, in that several SecY transmembrane helices are shifted to create a hydrophobic crevasse open to the cytoplasm. Disulfide mapping indicate that this "pre-open" state of the bacterial SecYE complex was induced by the Fab fragment and SecA, which in common bind to SecY residues at the tip of the cytoplasmic domain. Our disulfide crosslinking experiments revealed that some of these SecY residues contact specific residues of SecA that are otherwise buried and unavailable in the isolated SecA molecule. These results suggest that the motor components of the Sec machinery undergo cooperative conformational changes upon their interaction, presumably as important steps for the entrance of the preprotein-SecA complex and enhancement of the ATPase activity of SecA.
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[Journal Article] Conformational transition of Sec machinery inferred from bacterial SecYE structures.2008
Author(s)
Tsukazaki, T. *, Mori, H. *, Fukai, S., Ishitani, R., Mori, T., Dohmae, N., Perederina, A., Sugita, Y., Vassylyev, D.G., Ito, K., Nureki O.
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Journal Title
Nature 455
Pages: 988-991
Peer Reviewed
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[Presentation] Structural analysis of bacterial Sec translocation machinery.2009
Author(s)
Tsukazaki T., Mori H., Echizen, Y., Fukai, S., Ishitani, R., Mori, T., Dohmae, N., Perederina, A., Sugita, Y., Vassylyev, D.G., Ito, K., Nureki, O.
Organizer
International Symposium‘Innovative Nanoscience of Supermolecular Motor Proteins Working in Biomembranes'
Place of Presentation
Kyoto, Japan
Year and Date
20090908-20090910
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[Presentation] Functional dissection of bacterial Sec translocation machinery.2009
Author(s)
Mori H., Tsukazaki T., Echizen, Y., Ishitani, R., Nureki, O., Ito, K.
Organizer
International Symposium‘Innovative Nanoscience of Supermolecular Motor Proteins Working in Biomembranes'
Place of Presentation
Kyoto, Japan
Year and Date
20090908-20090910
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[Presentation] Crystal structure of T. thermophilus Sec translocon with an anti-SecY Fab fragment implies a translocation-initiating state of SecY2008
Author(s)
Tsukazaki T., Mori H., Fukai, S., Ishitani, R., Mori, T., Dohmae, N., Perederina, A., Sugita, Y., Vassylyev, D.G., Ito, K., Nureki, O.
Organizer
第31回日本分子生物学会年会、第81回日本生化学会大会合同大会シンポジウム「タンパク質機能発現システム-シャペロンからトランスロケーターまで」
Place of Presentation
神戸
Year and Date
20081209-20081212
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[Presentation] 細菌型Secトランスロコン複合体の結晶構造解析2008
Author(s)
Tsukazaki T., Mori H., Fukai, S., Ishitani, R., Perederina, A., Vassylyev, D.G., Ito, K., Nureki, O.
Organizer
第46回日本生物物理学会シンポジウム「膜を介した物質輸送・シグナル変換の構造基盤」
Place of Presentation
福岡
Year and Date
20081203-20081205
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