Isolation of drugs which rescue insulin resistance by activating GLUT4 transport activity.

分离通过激活 GLUT4 转运活性来缓解胰岛素抵抗的药物。

• 批准号: 23658222
• 负责人: SHIN-ICHIRO Takahashi
• 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 Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-23658222/
• 关键词: 糖輸送体(GLUT)4; 糖取り込み; インスリン; 成長ホルモン; インスリン抵抗性; 脂肪細胞; 糖輸送体（GLUT）4

We have reported that chronic GH pretreatment inhibited insulin-induced glucose uptake without affecting insulin-induced GLUT4 translocation. This study was undertaken to identify Akt substrates and GLUT4 postranslational modification that regulate GLUT4 transport activity. In addition, in order to rescue insulin resistance we searched for the small molecular chemicals and antibodies that modulate GLUT4 transport activity.At first, we succeeded to isolate the Akt substrates, AS47, whose insulin-inducedphosphorylation was suppressed by GH pretreatment. Knockdown of AS47 inhibited insulin-induced glucose uptake without affecting GLUT4 translocation, suggesting that AS47 plays important roles in GLUT4 transport activity.Next, we searched for posttranslational modification motif in a GLUT4 molecule. GLUT4 was site-directed mutagenized in some modification sites and these mutants were transfected into HEK293 cell. And then glucose uptake was measured. Mutation at the phosphorylation site of GLUT4 (GLUT4-P) significantly enhanced glucose transport activity, suggesting that phosphorylation impaired GLUT4 transport activity. Moreover, we have already succeeded to isolate antibodies, which recognize GLUT4 extracellular domain.In this study we succeeded to identify the Akt substrate and GLUT4 posttranslational modification, which play important roles in GLUT4 transport activity. Chemicals or antibodies, which interact with the Akt substrate and GLUT4 could be candidates tocure insulin resistance.

我们已经报道，长期 GH 预处理抑制胰岛素诱导的葡萄糖摄取，而不影响胰岛素诱导的 GLUT4 易位。本研究旨在鉴定调节 GLUT4 转运活性的 Akt 底物和 GLUT4 翻译后修饰。此外，为了挽救胰岛素抵抗，我们寻找调节GLUT4转运活性的小分子化学物质和抗体。首先，我们成功分离出Akt底物AS47，其胰岛素诱导的磷酸化被GH预处理抑制。 AS47的敲低抑制了胰岛素诱导的葡萄糖摄取，而不影响GLUT4易位，这表明AS47在GLUT4转运活性中发挥重要作用。接下来，我们在GLUT4分子中寻找翻译后修饰基序。对GLUT4的一些修饰位点进行定点诱变，并将这些突变体转染至HEK293细胞中。然后测量葡萄糖摄取量。 GLUT4 (GLUT4-P) 磷酸化位点的突变显着增强了葡萄糖转运活性，表明磷酸化损害了 GLUT4 转运活性。此外，我们已经成功分离出识别 GLUT4 胞外结构域的抗体。在本研究中，我们成功鉴定了 Akt 底物和 GLUT4 翻译后修饰，它们在 GLUT4 转运活性中发挥着重要作用。与 Akt 底物和 GLUT4 相互作用的化学物质或抗体可能是治疗胰岛素抵抗的候选药物。

项目成果

AP-1 complex regulates intracellular localization of insulin receptor substrate-1 required for insulin-like growth factor-I-dependent cell proliferation

AP-1 复合物调节胰岛素样生长因子 I 依赖性细胞增殖所需的胰岛素受体底物 1 的细胞内定位

• 发表时间: 2013
• 期刊: Mol. Cell. Biol.
• 作者: Yoneyama Y;Matsuo M;Take K;Kabuta T;Chida K;Hakuno F;Takahashi SI,
• 通讯作者: Takahashi SI,

動物細胞制御学研究室

动物细胞控制实验室

Diacylglycerol kinase ζ, negatively modulated GLUT4 translocation in 3T3-L1 adipocytes

二酰甘油激酶 ζ，负调节 3T3-L1 脂肪细胞中的 GLUT4 易位

• 发表时间: 2012
• 作者: Hakuno F;Ando Y;Kakino M;Fujimoto H;Chida K;Takahashi SI
• 通讯作者: Takahashi SI

The AP-1 Complex Regulates Intracellular Localization of Insulin Receptor Substrate 1, Which Is Required for Insulin-Like Growth Factor I-Dependent Cell Proliferation

• DOI: 10.1128/mcb.01394-12
• 发表时间: 2013-03
• 期刊: Molecular and Cellular Biology
• 影响因子: 5.3
• 作者: Yosuke Yoneyama;M. Matsuo;Kazumi Take;Tomohiro Kabuta;K. Chida;F. Hakuno;Shin-ichiro Takahashi

Diacylglycerol kinase ζ , a novel binding partner of insulin receptor substrates, negatively modulated GLUT4 translocation in 3T3-L1 adipocytes.

二酰基甘油激酶 z 是胰岛素受体底物的一种新型结合伴侣，可负调节 3T3-L1 脂肪细胞中的 GLUT4 易位。

• 发表时间: 2012
• 作者: Hakuno F;Ando Y;Kakino M;Fujimoto H;Chida K;Takahashi SI
• 通讯作者: Takahashi SI