Structural fluctuations of protein complexes measured by combination of protein engineering and solvent quenching meshod
| Project/Area Number |
23770194
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biophysics
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| Research Institution | Yamagata University (2012-2013)
National Institutes of Natural Sciences Okazaki Research Facilities (2011) |
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Principal Investigator |
MAKABE koki 山形大学, 理工学研究科, 准教授 (20508072)
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| Project Period (FY) |
2011 – 2013
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | タンパク質工学 / シャペロン / 水素交換 / GroEL / 蛋白質 / 構造揺らぎ / タンパク質 / 水素重水素交換 |
|---|
| Research Abstract |
In this project, we have been studying the structural fluctuations of E. coli chaperonin GroEL/ES system by using DMSO-quenched H/D exchange methods. For GroES H/D exchange experiments, we have successfully determined the exchange rates for 33 amide protons out of 94 amide protons in GroES. For the remaining 61 amide protons, we have determined the lower limit of the exchange rates. We found that only 10 amide protons have a protection factor larger than 10 to the power 6th. This result indicates that the free GroES is highly flexible in significant portions. From crystal structure analysis of N-terminal mutants of alpha-lactalbumin, a substrate protein of GroEL/ES system, we found the hydrogen bonding of the N-terminal residue is important for its stability.
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Report
(4 results)
Research Products
(32 results)
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[Journal Article] Dissection of the ATP-dependent conformational change cycle of a group II chaperonin.2014
Author(s)
Nakagawa A, Moriya K, Arita M, Yamamoto Y, Kitamura K, Ishiguro N, Kanzaki T, Oka T, Makabe K, Kuwajima K, Yohda M.
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Journal Title
J Mol Biol.
Volume: 426
Issue: 2
Pages: 447-459
DOI
Related Report
Peer Reviewed
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[Journal Article] ATP dependent rotational motion of group II chaperonin observed by X-ray single molecule tracking2013
Author(s)
Sekiguchi H, Nakagawa A, Moriya K, Makabe K, Ichiyanagi K, Nozawa S, Sato T, Adachi S, Kuwajima K, Yohda M, Sasaki YC
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Journal Title
Related Report
Peer Reviewed
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[Journal Article] ATP dependent rotational motion of group II chaperonin observed by X-ray single molecule tracking.2013
Author(s)
Sekiguchi H, Nakagawa A, Moriya K, Makabe K, Ichiyanagi K, Nozawa S, Sato T, Adachi S, Kuwajima K, Yohda M, Sasaki YC.
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Journal Title
PLoS One
Volume: 8
Issue: 5
Pages: e64176-e64176
DOI
Related Report
Peer Reviewed
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