Studies on Functions of bioactive phospholipids

生物活性磷脂的功能研究

• 批准号: 06304050
• 负责人: NOZAWA Yoshinori
• 金额: $ 17.66万
• 依托单位: GIFU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06304050/
• 关键词: Phospholipases (A2, C,D); Bioactive phospholipid; PAF-receptor; PAF-acetylhydrolase; GPI-anchor; PA phosphohydrolase; PIP2-binding protein; ホスホリパーゼA2,C,D; 情報変換素子; 情報変換ホスホリパーゼ; スフィンゴ脂質; 膜リン脂質

(1) PLD was synergistically activated by PKCalpha and RhoA.Rat PLD2 cDNA was cloned and mRNA levels of rPLD1a, 1b and 2 were observed to change during proliferation and differentiation (Nozawa). (2) cDNA of PAF-acetylhydrolase of guinea-pig blood plasma was isolated and the primary structure of the enzyme was determined (Nojima). (3) The CD14 was found to be the endotoxin (LPS) receptor. The signaling through LPS receptor requires PC breakdown by PLD (Nishijima). (4) The PLCdelta4 cDNA was cloned and its mRNA was expressed in the regenerating rat liver cell nuclei. A downstream molecules of Ash/Grb2 was PIP2 phosphatase (Takenawa). (5) DGKdelta and PAP2a, b were cloned. PAP2a was identified to be a Wunen homologue and PAP2b was Dri gene (Kanoh). (6) The image analysis with fluorescence microscope has shown that PS and some cytosolic factor(s) were required for the PS transfer to Golge bodies (Ohki). (7) PAF-acetylhydrolase was found to be composed of 3 subunits (alpha1, alpha2=catalytic subunits). beta subunit is the product of LIS-1 gene of Miller-Dieker syndrome (Inoue). (8) The amino acid sequences of the N-terminal and internal part of amino peptidase N were determined. GPI-anchored protein(s) was found in Sulfolobus (Ikezawa). (9) Expression of the intestinal phospholipase B-cDNA revealed that this enzyme was a novel serine esterase (Okamoto). (10) Sphingosine showed structural specific inhibition of DNA polymerase. PLA_2 purified from the chromatin fraction of rat ascitic hapatoma was 17kDa PLA_2II which was specifically localyzed (Koizumi). (11) It was demonstrated that 2 splicing variants of PAF-receptor genes were present and that leucocyte regulated by distinct promotors. Physiological functions were examined by using knock-out mice (Shimizu).

(1)通过PKCalpha和RhoA协同激活PLD。克隆大鼠PLD 2 cDNA，观察rPLD 1a、1b和2的mRNA水平在增殖和分化期间的变化（Nozawa）。(2)分离豚鼠血浆的PAF-乙酰水解酶的cDNA，并确定该酶的一级结构（Nojima）。(3)发现CD 14是内毒素（LPS）受体。通过LPS受体的信号传导需要PC被PLD（Nishijima）分解。(4)克隆了PLC δ 4 cDNA，并在再生大鼠肝细胞核中表达其mRNA。Ash/Grb 2的下游分子是PIP 2磷酸酶（Takenawa）。(5)克隆了DGKdelta和PAP 2a、B。PAP 2a被鉴定为Wunen同源基因，PAP 2b为Dri基因（Kanoh）。(6)荧光显微镜图像分析表明，PS向Golge小体（Ohki）的转移需要PS和一些胞质因子。(7)PAF-乙酰水解酶由3个亚基组成（α 1，α 2 =催化亚基）。β亚基是Miller-Dieker综合征（Inoue）LIS-1基因的产物。(8)测定了氨基肽酶N的N端和内部氨基酸序列。在硫化叶菌中发现了GPI锚定蛋白。(9)肠磷脂酶B-cDNA的表达表明，这种酶是一种新的丝氨酸酯酶（冈本）。(10)鞘氨醇对DNA聚合酶有结构特异性抑制作用。从大鼠腹水肝癌染色质中纯化的PLA_2为17kDaPLA_2 Ⅱ，经Koizumi法定位。(11)结果表明，PAF受体基因存在2种剪接变体，白细胞受不同的启动子调控。通过使用基因敲除小鼠（Shimizu）检查生理功能。

项目成果

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Watanabe T.: "Prostaglandin F2a stimulates formation of p21ras-GTP complex and mitogen-activated protein kinase in NIH-3T3 cells via Gq-protein-coupled pathway" J. Biol. Chem.270. 8984-8990 (1995)

Watanabe T.：“前列腺素 F2a 通过 Gq 蛋白偶联途径刺激 NIH-3T3 细胞中 p21ras-GTP 复合物和丝裂原激活蛋白激酶的形成”J. Biol。

K. Ouahchi: "Ataxia with isolated vitamin E deficiency is caused by mutations in the α-tocopherol transfer protein" Nature Genet.9. 141-145 (1995)

K. Ouahchi：“伴有维生素 E 缺乏的共济失调是由 α-生育酚转移蛋白突变引起的”Nature Genet.9（1995 年）。