The establishment of screening system to identify compoundsthat target BMP receptor-associated molecules

建立针对BMP受体相关分子的化合物筛选体系

基本信息

批准号：
07557373
负责人：
UENO Naoto
金额：
$ 1.15万
依托单位：
HOKKAIDO UNIVERSITY
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 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557373/
关键词：
BMP bone formation receptor Ser Thr kinase Signal transduction

项目摘要

In order to develop efficienct drugs that mimic the endogenous pathway of cytokines, precise molecular mechanism of signal transduction has to be clarified. Based on the mechanism, screening for compunds that modify the signaling pathway and thus act as agonist or antagonist can be achieved. TGF-beta activating kinase 1 (TAK1) and its activator, TAK1 binding protein 1 (TAB1) are implicated in TGF-beta and bone morphogenetic protein (BMP) signaling. However, the precise molecular mechanism by which ligand-ligated type I receptors induce TAB1 to activate TAK1 remains to be identified. To clarify BMP receptor mediated signaling, cytoplasmic interactors of BMP type Ia receptor (BMPRIa) were isolated with the use of a yeast two-hybrid system. One of the interactors isolated bound both TAB1 and BMPRIa in vivo. Sequence analysis revealed that this clone encoded a previously identified a denovirus E1A-associated protein, BS69. Although it contained E1A binding domain of BS69, NH2-terminal 12 amino acids of this clone were different from corresponding region of BS69. Therefore, we designated this molecule as BMP receptor associated molecule2 (BRAM2) and used in further experiments. The BRAM2 bound BMPRIa and TGF-beta type I receptor (TbetaRI) and augmented TAB1 binding to BMPRIa and TbetaRI in vivo. Furthermore, TAK1 and TAB1 mediated activation of plasminogen activator inhibitor-1 (PAI-1) gene promoter was strength ened by the expression of BRAM2. These results suggest that BRAM2 regulates the activity of TAB1 and is involved in TGF-beta and BMP signaling. Based on this knowledge, compunds that stimulate binding of BRAM2 to BMPR and may serve as agonists can now be screeened.

为了开发模拟细胞因子内源性途径的有效药物，必须阐明信号转导的精确分子机制。基于该机制，可以筛选修饰信号通路从而充当激动剂或拮抗剂的化合物。 TGF-β 激活激酶 1 (TAK1) 及其激活剂 TAK1 结合蛋白 1 (TAB1) 参与 TGF-β 和骨形态发生蛋白 (BMP) 信号传导。然而，配体连接的 I 型受体诱导 TAB1 激活 TAK1 的精确分子机制仍有待确定。为了阐明 BMP 受体介导的信号传导，使用酵母双杂交系统分离了 Ia 型 BMP 受体 (BMPRIa) 的细胞质相互作用因子。分离出的相互作用因子之一在体内结合 TAB1 和 BMPRIa。序列分析显示，该克隆编码先前鉴定的腺病毒 E1A 相关蛋白 BS69。虽然它含有BS69的E1A结合结构域，但该克隆的NH2末端12个氨基酸与BS69的相应区域不同。因此，我们将该分子命名为BMP受体相关分子2（BRAM2）并用于进一步的实验。 BRAM2 结合 BMPRIa 和 TGF-β I 型受体 (TbetaRI)，并在体内增强 TAB1 与 BMPRIa 和 TbetaRI 的结合。此外，BRAM2 的表达增强了 TAK1 和 TAB1 介导的纤溶酶原激活物抑制剂-1 (PAI-1) 基因启动子的激活。这些结果表明 BRAM2 调节 TAB1 的活性并参与 TGF-β 和 BMP 信号传导。基于这一知识，现在可以筛选刺激 BRAM2 与 BMPR 结合并可作为激动剂的化合物。