Attempt for regulating biologial activities by use of molecular recognition of SH2 and SH3 domains.

尝试通过使用SH2和SH3结构域的分子识别来调节生物活性。

• 批准号: 07558091
• 负责人: TAKENAWA Tadaomi
• 金额: $ 8.32万
• 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 (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07558091/
• 关键词: N-WASP; Ash; Grb2; filopodia; actin; tyrosine kinase; Grb2; SH2ドメイン; SH3ドメイン; ホスホリパーゼCγ1; SH_2ドメイン; SH_3ドメイン; ホスホリパーゼC_γ1

We found an adaptor protein, Ash/Grb2 which transmits tyrosine kinase signals to downstream, resulting in activation of Ras. This protein has one SH2 and two SH3 domains. In this project, we attempted to clarify binding proteins to Ash/Grb2 SH3 domain, and then purified SH3-binding proteins from bovine brain. Among them, two proteins (150 kDa and 65 kDa proteins) were new. After cDNA clonings of these proteins, 150 kDa protein was found to be one of synaptojamin isozymes. On the other hand, 65 kDa protein was found to be homologous to Wiskott-Aldrich syndrome protein (WASP). We therefore named it N-WASP.N-WASP has a PH domain, GBD/CRIB motif, IQ motif, verprolin-like motif, cofilin-like motif and proline-rich sequences. It binds to Ash/Grb2 SH3 domain and is activated downstream of tyrosine kinases. Activated N-WASP polymerizes actin filament in Cdc42 dependent manner, resulting in filopodia formation.

我们发现了一个衔接蛋白Ash/Grb 2，它将酪氨酸激酶信号传递到下游，导致Ras的激活。该蛋白具有一个SH 2和两个SH 3结构域。本研究试图从牛脑中分离出与Ash/Grb 2 SH 3结构域结合的蛋白，并纯化出与SH 3结构域结合的蛋白。其中150 kDa和65 kDa蛋白为新蛋白。对这些蛋白进行cDNA克隆后，发现150 kDa蛋白是突触素同工酶之一。另一方面，发现65 kDa蛋白与Wiskott-Aldrich综合征蛋白（WASP）同源。N-WASP具有PH结构域、GBD/CRIB基序、IQ基序、verprolin样基序、cofilin样基序和富含脯氨酸的序列。它与Ash/Grb 2 SH 3结构域结合，并在酪氨酸激酶下游被激活。活化的N-WASP以Cdc 42依赖的方式聚合肌动蛋白丝，导致丝状伪足形成。

项目成果

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K.Fukami、N.Sawada、T.Endo 和 T.Takenawa：“鸡骨骼肌 α-肌动蛋白中 PIP2 结合位点的鉴定”J.Biol.Chem.271。

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J.C.Hay等人：“Ca 2+ 激活分泌所需的ATP依赖性肌醇磷酸化”Nature。

M.Matsuda,et al: "Interaction between the amino-terminal SH3 domain of Crk and its natural target proteins" J.Biol.Chem.271. 14468-14472 (1996)

M.Matsuda 等人：“Crk 氨基末端 SH3 结构域与其天然靶蛋白之间的相互作用”J.Biol.Chem.271。

H.Yu, et al.: "Phosphatidylinosito 4,5-bisphosphate reverses the inhibition of RNA transcription caused by histon Hl" Eur.J.Biochem.(in press).

H.Yu 等人：“磷脂酰肌醇 4,5-二磷酸逆转组蛋白 H1 引起的 RNA 转录抑制”Eur.J.Biochem.（出版中）。

M.Fukuoka, et al.: "Identification of N-WASP homologs in human and rat brain" Gene. 196. 43-48 (1997)

M.Fukuoka 等人：“人和大鼠大脑中 N-WASP 同源物的鉴定”基因。