| Project/Area Number |
11460039
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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 |
応用微生物学・応用生物化学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
MURATA Kousaku Kyoto University, Agriculture, Professor, 農学研究科, 教授 (90142299)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAI Shigeyuki Kyoto University, Agriculture, Research Associate, 農学研究科, 助手 (00303909)
HASHIMOTO Wataru Kyoto University, Agriculture, Associate Professor, 農学研究科, 助教授 (30273519)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥14,300,000 (Direct Cost: ¥14,300,000)
Fiscal Year 2001: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1999: ¥6,600,000 (Direct Cost: ¥6,600,000)
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| Keywords | sphingomonas / alginate lyase / autoprocessing / protease / x-ray crystallography / β-elimination mechanism / 遺伝情報多重性 / プロテオーム / 遺伝情報量 / 多触媒中心酵素 / 翻訳後修飾 / X線結晶構造解析 / アルギン酸 / Shingomonas属細菌 / プロテオーム解析 |
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| Research Abstract |
The intracellular alginate lyases I,II, and III are coded by one gene and produced through sequential post-translational reactions from a common pre-cursor protein Po : Po → I + 5 kDa peptide, I → II +III. The protease-like activity of I is completely inhibited by EDTA or O- phnonthroline, thus indicates that the I is a multi- catalytic site enzyme currying II, III, and protease. The protease site on I is thought to be created through the excision of N-terminal 5 kDa peptide from Po, since Po has no protease activity. An important biological problem is then raised as to how the protease site is generated and how / where the information is described in the base and amino acid sequences. The clarification of these addresses may lead to the postulation of a novel theory concerning the abundance of information contained in a gene. To solve the problems, the knowleges on the structure / function relationship of alginate lyases I, II, and III, as well as their precursor protein Po. Among these, the structure / function of III was completely analyzed through X-ray crystallography and detailed lyase reaction mechanism was clarified. Especially, a novel β-elimination reaction mechanism involving Tyr residue, but not His, was constructed with an emphasis of Low-Barrier Hydrogen Bond Theory.
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