High-resolution Neutron Structural Analysis of Two States of a Bilin Reductase PcyA
| Project/Area Number |
24570122
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Ibaraki University |
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Principal Investigator |
UNNO Masaki 茨城大学, 理工学研究科, 教授 (10359549)
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| Co-Investigator(Renkei-kenkyūsha) |
FUKUYAMA Keiichi 大阪大学, 大学院工学研究科, 招へい研究員 (80032283)
WADA Kei 宮崎大学, テニュアトラック推進機構, テニュアトラック准教授 (80379304)
TAMADA Taro 原子力機構, 量子ビーム, グループリーダー (50391248)
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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 |
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | 中性子結晶構造解析 / 酵素 / 水素 / 還元反応 / フィコシアノビリン / コンフォメーション / 還元 / 色素 / 光合成 / 水素原子 / ヒドロニウムイオン / J-PARC / ビリン還元酵素 / 反応機構 / 光合成色素 / 光反応 / 中性子結晶解析 / 回折 / 光応答 / プロトネーション |
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| Outline of Final Research Achievements |
PcyA catalyzes two of the steps of two-proton-coupled two-electron reduction of biliverdin (BV）, to phycocyanobilin. Revealing the protonation states in the active site of PcyA and the substrate BV is important for understanding this unique reaction mechanism. In the present study, we determined the neutron crystal structure of PcyA in complex with BV. The structure revealed the protonation states of BV and the surrounding residues. We found that two forms of BV, neutral BV and protonated BVH+, were coupled with the two conformation/protonation states of the essential residue Asp105. Further, His88 and His74 near BV were singly protonated and were connected with an intervening hydronium ion. To our knowledge, this is the third example of a hydronium ion in a protein structure. Neutron analysis also revealed how X-ray irradiation of the PcyA-BV crystal altered the structure of the PcyA-BV complex.
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Report
(4 results)
Research Products
(18 results)
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[Journal Article] Insights into the Proton Transfer Mechanism of a Bilin Reductase PcyA Following Neutron Crystallography.2015
Author(s)
Unno, M., Ishikawa-Suto, K., Kusaka, K., Tamada, T., Hagiwara, Y., Sugishima, M., Wada, K., Yamada, T., Tomoyori, K., Hosoya, T., Tanaka, I., Niimura, N, Kuroki, R., Inaka, K., Ishihara, M., Fukuyama, K.
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Journal Title
J. Am. Chem. Soc.
Volume: in press
Issue: 16
Pages: 5452-5460
DOI
Related Report
Peer Reviewed / Acknowledgement Compliant
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[Presentation] Neutron Structure of Phycocyanobilin:oxidoreductase Complexed with Biliverdin2014
Author(s)
Masaki Unno, Kumiko Ishikawa-Sudo, Katsuhiro Kusaka, Taro Tamada, Yoshinori Hagiwara, Masakazu Sugisima, Kei Wada, Taro Yamada, Katsuaki Tomoyori, Takaaki Hosoya, Ichiro Tanaka, Nobuo Niimura, Ryota Kuroki, Koji Inaka, Makiko Ishihara, and Keiichi Fukuyama
Organizer
23rd Congress and General Assembly of the International Union of Crystallography
Place of Presentation
Montreal
Year and Date
2014-08-09
Related Report
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