Analysis of the mechanism of activation and co-translational insertion of an essential trace element, selenium, into polypeptide
| Project/Area Number |
15370043
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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 |
Structural biochemistry
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
ESAKI Nobuyoshi KYOTO UNIVERSITY, Institute for Chemical Research, Professor, 化学研究所, 教授 (50135597)
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| Co-Investigator(Kenkyū-buntansha) |
KURIHARA Tatsuo KYOTO UNIVERSITY, Institute for Chemical Research, Associate Professor, 化学研究所, 助教授 (70243087)
MIHARA Hisaaki KYOTO UNIVERSITY, Institute for Chemical Research, Assistant Professor, 化学研究所, 助手 (30324693)
YOSHIMURA Tohru Nagoya University, Graduate School of Bioagricultural Science, 大学院・生命農学研究科, 教授 (70182821)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥15,400,000 (Direct Cost: ¥15,400,000)
Fiscal Year 2004: ¥5,900,000 (Direct Cost: ¥5,900,000)
Fiscal Year 2003: ¥9,500,000 (Direct Cost: ¥9,500,000)
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| Keywords | selenium / selenocysteine lyase / selenoprotein / phytoremediation / RNAi / x-ray crystal structural analysis / 培養細胞 / HeLa / 脳 / セレン欠乏 |
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| Research Abstract |
Selenium, an essential trace element, is incorporated into a specific position of selenoprotein in the form of selenocysteine residues and plays a various important roles in organisms. In this study, we investigated the function of proteins involved in the metabolism of selenium.
1.Selenocysteine lyase (SCL) catalyzes the decomposition of L-selenocysteine into L-alanine and selenium. We employed the yeast two-hybrid system using mouse SCL as a bait to search for a protein(s) that interacts with SCL. We identified major urinary protein (MUP) as a potential partner of SCL. Our analysis revealed that SCL inhibits the interaction between MUP and 2-naphtol in vitro.
2.The mouse SCL gene was expressed in the cytosol or chloroplast of Arabidopsis thaliana. The selenium-tolerance of the plant was greatly improved, indicating that SCL is useful for phyroremediation.
3.The distribution of selenium and selenoprotein mRNAs in moue brain were determined by quantification of selenium and quantitative RT-PCR, respectively.
4.siRNAs targeted to SCL were transfected with human HeLa cells and mouse Al cells to knock-down the expression of SCL. We found that siRNAs against SCL decreases the expression of selenoproteins in both cells. The results suggest that SCL is involved in the biosynthesis of selenoprotein.
5.Rat SCL was crystallized and its three-dimensional structure was solved. The structure revealed that SCL has a catalytically essential cysteine residue on a extended lobe.
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Report
(3 results)
Research Products
(19 results)
