Elucidation of mechanisms for catalytic center maturation and catalysis of nitrile hydratase family enzyme
| Project/Area Number |
21350089
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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 |
Chemistry related to living body
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
ODAKA Masafumi 東京農工大学, 大学院・工学研究院, 准教授 (20224248)
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| Co-Investigator(Renkei-kenkyūsha) |
KUROKI Ryota 独立行政法人日本原子力研究開発機構, 量子ビーム応用研究部門, ユニット長 (30391246)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000)
Fiscal Year 2011: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2010: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2009: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
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| Keywords | 水和反応 / 加水分解 / 酵素反応 / 結晶構造解析 / 時間分割構造解析 / 翻訳後修飾 / タンパク質成熟化 / アクセサリータンパク質 / 構造生物学 / 中性子 / 生物・生体工学 / バイオリアクター / X線結晶構造解析 |
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| Research Abstract |
By performing time-resolved X-ray crystallography of Fe-type nitrile hydratase(NHase) whoseβArg56 were substituted by Lys, we obtained the structure of the enzyme-substrate complex. By replacing two Arg residues resided in the active site pocket of a NHase family enzyme, thiocyanate hydrolase(SCNase), we showed that the size as well as the surface charge of the substrate binding pocket was important for the substrate specificities of SCNase. As for the maturation mechanism, we found that the SCNase activator protein, P15K, is responsible for the refolding, metal-insertion and post-translational modification of Cys ligand in the catalytic subunit,γ. The metal-incorporatedγsubunit form the complex with the other subunits,αandβ, to form the mature SCNase.
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Report
(4 results)
Research Products
(51 results)
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[Presentation] Catalytic mechanism of nitrile hydratase proposed by time-resolved X-ray crystallography using a novel substrate, tert-butylisonitrile2009
Author(s)
Odaka M., Hashimoto, K., Suzuki, H., Taniguchi, K., Noguchi, T., Yohda, M.
Organizer
The 14th International Conference on Biological Inorganic Chemistry(ICBIC14)
Place of Presentation
Nagoya, Japan
Year and Date
2009-07-30
Related Report
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