Functional analysis of phosphatidylinositol 5-kinases (PIP5K)

磷脂酰肌醇 5-激酶 (PIP5K) 的功能分析

• 批准号: 11680624
• 负责人: SHIBASAKI Yoshikazu
• 金额: $ 2.5万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680624/
• 关键词: inositol; kinase; actin; low molecular weight G protein; membrane traffic; vesicular transport; cytoskeleton; キナーゼ; ホスホリパーゼD; PHドメイン; ラフリング

(1) Analysis of mutant PIP5K : By deleting either N-terminus or C-terminus, the kinase activity of PIP5K was localized in central 380 amino acids region.(2) Intracellular localization of PIP5K : Endogenous PIP5K was hard to detect as the expression was very low. Exogenous PIP5K was found mainly on the cytoplasmic surface of the plasma membrane and also in the cytoplasm.(3) Products of PIP5K reaction : In addition to PI (4,5) P2, small amount of PI (5) P and PI (3,5) P2 was produced.(4) Actin polymerization : By overexpression of PIP5K, actin polymerization was greatly enhanced.(5) Interactin between Rac and PIP5K : In platelet semiintact system, Rac and PIP5K are both involved in thrombin stimulated actin polymerization process. When a Rac mutant was introduced, the polymerization was inhibited, suggesting that the interaction between Rac and PIP5K is necessary for actin polymerization

(1)突变体PIP 5 K的分析：通过缺失N-末端或C-末端，PIP 5 K的激酶活性定位于中心380个氨基酸区域。(2)PIP 5 K的细胞内定位：内源性PIP 5 K很难检测，因为表达非常低。外源性PIP 5 K主要存在于质膜的胞质表面，也存在于细胞质中。(3)PIP 5 K反应产物：除PI（4，5）P2外，还产生少量PI（5）P和PI（3，5）P2。(4)肌动蛋白聚合：通过过表达PIP 5 K，肌动蛋白聚合大大增强。(5)Rac与PIP 5 K相互作用：在血小板半完整系统中，Rac和PIP 5 K均参与凝血酶刺激的肌动蛋白聚合过程。当引入Rac突变体时，聚合被抑制，表明Rac和PIP 5 K之间的相互作用是肌动蛋白聚合所必需的

项目成果

柴崎 芳一: "ホスファテジルイノシトール 5-キナーゼ"生 化 学. 71(3). 193-197 (1999)

Hoichi Shibasaki：“磷酸肌醇 5-激酶”生物化学 71(3)。

Tolias, KF: "Type Iα phosphatidyl-4-phosphate 5-kinase mediates Rac-dependent actin assembly"Current Biology. 10. 153-156 (2000)

Tolias，KF：“Iα 型磷脂酰 4 磷酸 5 激酶介导 Rac 依赖性肌动蛋白组装”《当代生物学》10. 153-156 (2000)。

Itoh, T: "Autophosphorylation of type I phosphatidyl phosphate Uinaseregulates its lipid Kinase activity"J. Biol. Chem.. 275. 19389-19394 (2000)

Itoh，T：“I 型磷脂酰磷酸酶的自磷酸化调节其脂质激酶活性”J。

Tolias K.F.: "Type I Phosphatidylinositol〜"J.Biol.Chem. 273. 18040-18046 (1998)

Tolias K.F.：“I 型磷脂酰肌醇 ~”J.Biol.Chem. 273. 18040-18046 (1998)

Tolias,KF: "Type I αphosphatidylinositol-4-phosphate 5-kinase mediates Rac-dependent actin assembly "Current Biology. 10. 153-156 (2000)

Tolias，KF：“I 型 α 磷脂酰肌醇-4-磷酸 5-激酶介导 Rac 依赖性肌动蛋白组装”《当代生物学》10. 153-156 (2000)。