2013 Fiscal Year Final Research Report
Structural basis of ATP hydrolysis mechanism of V1-ATPase motor
| Project/Area Number |
23370047
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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 |
Structural biochemistry
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| Research Institution | Chiba University |
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Principal Investigator |
MURATA Takeshi 千葉大学, 理学(系)研究科(研究院), 准教授 (80415322)
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| Co-Investigator(Kenkyū-buntansha) |
YAMATO Ichiro 東京理科大学, 基礎工学部, 教授 (70111458)
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| Co-Investigator(Renkei-kenkyūsha) |
SAIJO Shinya 東京理科大学, 基礎工学部, 助教 (80419001)
MIZUTANI Kenji 東京理科大学, 基礎工学部, 博士研究員 (10525570)
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| Project Period (FY) |
2011-04-01 – 2014-03-31
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| Keywords | 分子モーター / 酵素反応 / V-ATPase / X線結晶構造解析 / ATP |
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| Research Abstract |
Vacuolar ATPases (V-ATPases) function as proton pumps in various membrane systems within the cells, which are involved in a number of processes such as bone resorption and cancer metastasis. The hydrophilic V1 portion is known as a rotary motor, in which a central axis DF complex rotates inside a hexagonally arranged catalytic A3B3 complex using ATP hydrolysis energy, although the molecular mechanism is not well defined due to a lack of high-resolution structural information.
We have established the in vitro expression, purification, and reconstitution of Enterococcus hirae V1-ATPase from the A3B3 and DF complexes. In this study, we solved the crystal structures of AMP-PNP-bound and Pi-bound V1-ATPase, and also characterized the several site-directed mutants. On the basis of these findings, we proposed the molecular mechanism of the rotary motor.
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[Journal Article] Mutant LV476-7AA of A-subunit of Enterococcus hirae V 1-ATPase : High affinity of A3B3 complex to DF axis and low ATPase activity2013
Author(s)
Md. Jahangir Alam, Ichiro Yamato, Satoshi Arai, Shinya Saijo, Kenji Mizutani, Yoshiko Ishizuka-Katsura, Noboru Ohsawa, Takaho Terada, Mikako Shirouzu, Shigeyuki Yokoyama, So Iwata, Yoshimi Kakinuma and Takeshi Murata
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Journal Title
SpringerPlus
Volume: 2
Pages: 689
Peer Reviewed
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[Journal Article] Basic properties of rotary dynamics of the molecular motor Enterococcus hirae V 1-ATPase2013
Author(s)
Yoshihiro Minagawa, Hiroshi Ueno, Mayu Hara, Yoshiko Ishizuka-Katsura, Noboru Ohsawa, Takaho Terada, Mikako Shirouzu, Shigeyuki Yokoyama, Ichiro Yamato, Eiro Muneyuki, Hiroyuki Noji, Takeshi Murata and Ryota Iino
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Journal Title
J. Biol. Chem
Volume: 288
Pages: 32700-32707
DOI
Peer Reviewed
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[Journal Article] Loose Binding of the DF Axis with the A 3B3 Complex Stimulates the Initial Activity of Enterococcus hirae V 1-ATPase2013
Author(s)
Md. Jahangir Alam, Satoshi Arai, Shinya Saijo, Kano Suzuki, Kenji Mizutani, Yoshiko Ishizuka-Katsura, Noboru Ohsawa, Takaho Terada, Mikako Shirouzu, Shigeyuki Yokoyama, So Iwata, Yoshimi Kakinuma, Ichiro Yamato and Takeshi Murata
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Journal Title
PLoS ONE
Volume: 8
Pages: e74291
DOI
Peer Reviewed
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[Journal Article] Rotation mechanism of Enterococcus hirae V 1-ATPase based on asymmetric crystal structures2013
Author(s)
Satoshi Arai, Shinya Saijo, Kano Suzuki, Kenji Mizutani, Yoshimi Kakinuma, Yoshiko Ishizuka-Katsura, Noboru, Ohsawa, Takaho Terada, Mikako Shirouzu, Shigeyuki Yokoyama, So Iwata, Ichiro Yamato and Takeshi Murata
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Journal Title
Nature
Volume: 493
Pages: 703-707
DOI
Peer Reviewed
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[Journal Article] Crystal structure of the central axis DF complex of V-ATPase2011
Author(s)
Shinya Saijo, Satoshi Arai, Khandoker M. M. Hossain, Kano Suzuki, Ichiro Yamato, Yoshimi Kakinuma, Yoshiko Ishizuka-Katsura, Noboru, Ohsawa, Takaho Terada, Mikako Shirouzu, Shigeyuki Yokoyama, So Iwata, and Takeshi Murata
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Journal Title
Proc. Natl. Acad. Sci. U. S. A.
Volume: 108
Pages: 19955-19960
DOI
Peer Reviewed
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