| Project/Area Number |
63570137
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | National Cardiovascular Center Research Institute |
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Principal Investigator |
MIYAKE Yoshihiro National Cardiovascular Center Research Institute ; Department of Biochemistry ; Head, 生化学部, 部長 (40028353)
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| Co-Investigator(Kenkyū-buntansha) |
MIYANO Motoshige National Cardiovascular Center Research Institute ; Department of Biochemistry ;, 生化学部, 研究員 (40166187)
INOUE Hiroyasu National Cardiovascular Center Research Institute ; Department of Biochemistry ;, 生化学部, 研究員 (40183743)
MOMOI Kyoko National Cardiovascular Center Research Institute ; Department of Biochemistry ;, 生化学部, 研究員 (90174359)
FUKUI Kiyoshi National Cardiovascular Center Research Institute ; Department of Biochemistry ;, 生化学部, 室長 (00175564)
渡辺 房男 国立循環器病センター研究所, 生化学部, 研究員 (40183719)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1988: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | D-amino acid oxidase / Renin-binding protein / Complementary DNA / Regulation of gene expression / Site-directed mutagenesis / Leucine zipper structure / Recombinant protein expression / Recombinant mutant proteins / 相補的DNA塩基配列 / 遺伝子発現 / 翻訳調節 / 変異体蛋白質精製 / 変異体蛋白質機能異常 / 遺伝子構造解析 / 大量発現と精製 / 遺伝子発現機構 / 部位特異的突然変異誘導体 / D-アミノ酸オキシダーゼ / 部位特異的変異誘導 / DNAライブラリー / 発現の組織特異性 |
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| Research Abstract |
1. A complementary DNA (cDNA) encoding human kidney D-amino acid oxidase (DAO) was isolated from a human kidney cDNA library, and the nucleotide sequence was determined.
2. A recombinant porcine DAO Was expressed in a large scale in. The expressed recombinant DAO was purified from the cell extract, and the molecular and enzymological properties were characterized.
3. Two kinds of recombinant mutant DAOs, in which Tyr-228 or His-307 is replaced with Phe or Leu, respectively, were expressed in Escherichia coli cells. The mutant DAOs were purified from the cell extract, and the functional abnormalities were shown.
4. A cDNA encoding rabbit kidney DAO was isolated from a rabbit kidney cDNA library, and the nucleotide sequence was determined. Moreover, the presence of translational suppression in the process of rabbit DAO gene expression was found.
5. A cDNA encoding mouse kidney DAO was isolated from a mouse kidney cDNA library, and the nucleotide sequence was determined.
6. cDNAs encoding porcine, human and rat renin-binding proteins (RnBP) were isolated from cDNA libraries and the nucleotide sequences were determined.
7. It was shown that RnBP is a new protein that has not been identified and it contains a new type of leucine zipper structure.
8. The leucine zipper motif in RnBP molecule was shown to participate in the protein function.
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