Regulatory mechanisms of multi-functional sheddase

多功能脱落酶的调控机制

• 批准号: 15570121
• 负责人: SAKANE Fumio
• 金额: $ 2.05万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15570121/
• 关键词: Diacylglycerol kinas; Phosphatidic acid; Osmotic shock; Phorbol ester; Phosphorylation; Oligomer; SAM; Pleckstrin homology; TACE; ホルボールエスチル; ジアシルグリセロール; 腫瘍壊死因子α; トランスロケーション; プロテインキナーゼC

We had ahrady obtained several lit s of evidence suggesting chat diacylglycerol kinase (DGK) 6 participates m the regulation of a multi-functional sheddase, tumor necrosis α-converning enzyme (TACE). As a next step, we by modification of DGK6 and its closely related isofoms (DGKs η and κ) in stimulated cells. DGKδ1 or its pleckstrin homology (PH) domain alone has been shown to be translocated to the plasma membranes from the cytoplasm in phorbol myristate acetate (PMA)-treated cells. In the present work we identified Ser-22 and Ser-26 within the PH domain as the PMA-and epidermal growth factor-dependent phosphorylation sites of DGKδ1. Experiments in vitro and with intact cells suggested that the conventional protein kinase C (cPKC) directly phosphorylated these Ser residues. Interestingly, the Asp-mutation, which mimics phosphoserine, at Ser-22 or Ser-26, markedly inhibited the translocation of full-length DGKδ1 and the PH domain, suggesting that the phosphorylation regulates negatively the enzyme translocation.We identified another DGKη isoform (η2, 135 kDa) that shared the same sequence with DGKη1 except for a sterile a motif (SAM) domain added at the C-terminus. DGKη2 was shown to form through its SAM domain homo-oligomers as well as hetero-oligomers with other SAM-containing DGKs (81 and 62). Interestingly, DGKη1 and DGKη2 were rapidly translocated from the cytoplasm to endosomes in response to stress stimuli. In this case, DGKη1 l was rapidly relocated back to the cytoplasm upon removal of stress stimuli, whereas DGKη2 exhibited sustained endosomal association.Recently, we identified a tenth member of the DGK family designated DGKκ. The new DGK isozyme has additionally 33 tandem repeats of Glu-Pro-Ala-Pro at the N-terminus. Interestingly, DGKκ, but not other type II DGKs, was specifically tyrosine-phosphorylated by Src-family kinase in H_2O_2-treated cells.

我们有Ahrady获得了几种证据，表明聊天二酰基甘油激酶（DGK）6参与了M的多官能sheddase，肿瘤坏死α-连接酶（TACE）的调节。作为下一步，我们通过修饰DGK6及其在刺激细胞中密切相关的异卵泡（DGKS和κ）。 DGKΔ1或其pleckstrin同源（pH）结构域已被证明被证明是易位的醋酸肉豆蔻酸酯（PMA）处理的细胞中细胞质中的质膜。在目前的工作中，我们将pH结构域内的SER-22和SER-26确定为DGKΔ1的PMA和表皮生长因子依赖性磷酸化位点。体外和完整细胞的实验表明，常规蛋白激酶C（CPKC）直接磷酸化了这些SER的保留。 Interestingly, the Asp-mutation, which mimics phosphoserine, at Ser-22 or Ser-26, markedly inhibited the translation of full-length DGKδ1 and the PH domain, suggesting that the phosphorylation regulates negatively the enzyme translocation.We identified another DGKη isoform (η2, 135 kDa) that shared the same sequence with DGKη1 except for a sterile a motif (SAM)在C末端添加的域。显示DGKη2通过其SAM域Homo-Oligomers以及其他含Sam的DGK（81和62）形成。有趣的是，DGKη1和DGKη2因应激刺激而迅速从细胞质转移到内体。在这种情况下，DGKη1L在去除应激刺激后迅速重新恢复到细胞质，而DGKη2表现出持续的内体关联。新的DGK同工酶在N末端还具有33个Glu-Pro-Ala-Pro的串联重复序列。有趣的是，DGKκ但没有其他II型DGK，是由SRC-家庭激酶在H_2O_2处理的细胞中特异性酪氨酸磷酸化的。

项目成果

生物薬科学実験講座 情報伝達物質[II](石橋貞彦, 市川厚, 堅田利明編)

生物制药科学实验课程信息传递物质[II]（石桥定彦、市川厚、片田俊明编）

• 作者: 坂根郁夫;山田恵子;加納英雄(分担執筆)
• 通讯作者: 加納英雄(分担執筆)

The plasma membrane translocation of diacylglycerol kinase δ1 is negatively regulated by conventional protein kinase C-dependent phosphorylation at Ser-22 and Ser-26 within the pleckstrin homology domain

• DOI: 10.1042/bj20040681
• 发表时间: 2004-09-15
• 期刊: BIOCHEMICAL JOURNAL
• 影响因子: 4.1
• 作者: Imai, S;Kai, M;Sakane, F
• 通讯作者: Sakane, F

Jia, Y-J., Kai, M., Wada, I., Sakane, F., Kanoh, H.: "Differential localization of lipid phosphate phosphatases 1 and 3 to cell surface subdomains in polarized MDCK cells"FEBS Lett.. 552(2-3). 240-246 (2003)

Jia, Y-J.、Kai, M.、Wada, I.、Sakane, F.、Kanoh, H.：“脂质磷酸磷酸酶 1 和 3 在极化 MDCK 细胞中细胞表面子结构域的差异定位”FEBS Lett.. 552（

Gene expression, cellular localization and enzymatic activity of diacylglycerol kinase isozymes in rat ovary and placenta

大鼠卵巢和胎盘中二酰甘油激酶同工酶的基因表达、细胞定位和酶活性

• 期刊: Cell Tissue Res. (in press)
• 作者: Toya;M.;Hozumi;Y.;Ito;T.;Takeda;M.;Sakane;F.;Kanoh;H.;Saito;H.;Hiroi;M.;Kurachi;H.;Kondo;H.;Goto;K.
• 通讯作者: K.

Expression and localization of diacylglycerol kinase isozymes and enzymatic features in rat lung.

二酰甘油激酶同工酶的表达和定位以及大鼠肺中的酶学特征。

• 期刊: Am.J.Physiol.Lung Cell Mol.Physiol. (in press)
• 作者: Katagiri;Y.;Ito;T.;Saino-Saito;S.;Hozumi;Y.;Suwabe;A.;Otake;K.;Sata;M.;Kondo;H.;Sakane;F.;Kanoh;H.;Kubota;I.;Goto;K.
• 通讯作者: K.