Single-molecule analysis of 1-nm stepping motion of linear motor carbohydrate hydrolase
| Project/Area Number |
24370062
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Partial Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Okazaki Research Facilities, National Institutes of Natural Sciences (2014)
The University of Tokyo (2012-2013) |
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Principal Investigator |
IINO RYOTA 大学共同利用機関法人自然科学研究機構(岡崎共通研究施設), 岡崎統合バイオサイエンスセンター, 教授 (70403003)
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| Co-Investigator(Kenkyū-buntansha) |
UCHIHASHI Takayuki 金沢大学, 数物科学系, 准教授 (30326300)
IGARASHI Kiyohiko 東京大学, 農学生命科学研究科, 准教授 (80345181)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000)
Fiscal Year 2014: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2013: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2012: ¥9,750,000 (Direct Cost: ¥7,500,000、Indirect Cost: ¥2,250,000)
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| Keywords | 分子モーター / １分子計測 / セルラーゼ / キチナーゼ / 1分子計測・操作 / モータータンパク質 |
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| Outline of Final Research Achievements |
In this study, using single-molecule fluorescence microscopy and high speed atomic force microscopy, we determined kinetic constants of the elementary reaction steps for carbohydrate hydrolase TrCel7A against cellulose Iα and IIII. TrCel7A displayed similar binding and dissociation rate constants for cellulose Iα and IIII and similar fractions of productive binding on cellulose Iα and IIII. Furthermore, once productively bound, TrCel7A processively hydrolyzes and moves along cellulose Iα and IIII with similar translational rates. With structural models of cellulose Iα and IIII, we propose that different susceptibilities at high TrCel7A concentration arise from surface properties of substrate, including ratio of hydrophobic surface and number of available lanes.
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Report
(4 results)
Research Products
(26 results)
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[Journal Article] Single-molecule Imaging Analysis of Elementary Reaction Steps of Trichoderma reesei Cellobiohydrolase I (Cel7A) Hydrolyzing Crystalline Cellulose Iα and IIII2014
Author(s)
Y.Shibafuji, A.Nakamura, T.Uchihashi, N.Sugimoto, S.Fukuda, H.Watanabe, M.Samejima, T.Ando, H.Noji, A.Koivula, K.Igarashi, R.Iino
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Journal Title
Journal of Biological Chemistry
Volume: 289 (29)
Issue: 20
Pages: 1405614065-1405614065
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article]2014
Author(s)
T.Ikeda, T.Tsukahara, R.Iino, M.Takeuchi, H.Noji
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Journal Title
Angew. Chem. Int. Ed.
Volume: 53, (accepted)
Issue: 38
Pages: 10082-10085
DOI
Related Report
Peer Reviewed
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