| Project/Area Number |
09480145
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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 |
Bioorganic chemistry
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| Research Institution | Kwansei Gakuin University |
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Principal Investigator |
KATSUMURA Shigeo Kwansei Gakuin University, School of Science, Professor, 理学部, 教授 (70047364)
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| Co-Investigator(Kenkyū-buntansha) |
IKEDA Kiyoshi Osaka University of Pharmaceutical Sciences, Department of Biochemistry, Professor, 薬学部, 教授 (50001053)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1997: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Keywords | phospholipase AィイD22ィエD2 / sphingomyelinase / synthesis of enzyme inhibitors / elucidation of inhibitory mechanism / interfacial recognition site / oxazolidinone substrate analog / synthesis of sphingomyelin / sphingolipid analog inhibitor / スフィンゴミエリナーゼ阻害剤 / メチレン気質類緑体 / 立体選択的合成 / ホフマン転位 / スフィンゴミエリン / 第2世代拮抗的阻害剤 / 2置換オキサゾリジノンリン脂酸 / 立体選択的オキサゾリジノン合成 / アミド型阻害剤とCa^<2+>との結合常数 / pH依存性 / アルデヒドテルペノイドの阻害機構 / ホスホリパーゼA_2阻害剤 / 3-(E)-メトキシカルボニルトリエナ-ル / リジンの選択的修飾 / ジヒドロピリジンの形成 / スフィンゴエミリナーゼ基質 / スフィンゴシンの立体選択的合成 |
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| Research Abstract |
Phospholipids are classified into glycerophospholipids and sphingophospholipids. Phospholipase AィイD22ィエD2 (PLAィイD22ィエD2) catalyzes the hydrolysis of the ester linkage at the sn-2 position of glycerophospholipids and responsibles for the metabolism of unsaturated fatty acids. Meanwhile, sphingomyelinase is the general term of the enzyme which catalyzes the hydrolysis of the phosphoester linkage of sphingomyelin to produce ceramide, which has been known as a lipid second messenger in mammalian cell membranes.
We found that 3-methoxycarbonyl-2, 4, 6-trienal (A) strongly inactivated phospholipase AィイD22ィエD2 which was isolated from bovine pancreas. The inactivation mechanism of PLAィイD22ィエD2 by the compound A was elucidated by means of the stereocontrolled synthesis of the derivatives of A, structure activity relationship, amino acid analysis of the complex with PLAィイD22ィエD2, and model reactions with amines. Thus, the compound A was found to selectively modify Lys-56 which is included in the
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interfacial recognition site of bovine pancreatic PLAィイD22ィエD2, by the MALDI-TOF-Mass peptide mapping analyses of the complex.
We also synthesized the new cyclic amide analog as the second-generation inhibitor having an oxazolidinone ring, because we had previously synthesized the oxazolidinone phospholipid analog which competitively acted on the catalytic site of PLAィイD22ィエD2.
On the other hand, in the study of sphingomyelinase, we achieved the efficient synthesis of (D)-erythro-sphigomyelin, and found that (D)-erythro stereochemistry is essentially important as the substrate for sphingomyelinase and the double bond in the backbone skeleton was not essential for the hydrolysis by B. cereus sphingomyelinase. Furthermore, we established the stereocontrolled efficient synthesis of sphingomyelin methylene and ethylene analogs, which are the first substrate-analog inhibitors, and they actually inhibited the hydrolytic ability of the enzyme. These analog inhibitors might be useful for the elucidation of the sphingomyelinase hydrolytic mechanism. Less
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