权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Structural studies on signal transduction related to Ash/Grb2

Ash/Grb2相关信号转导的结构研究

基本信息

批准号：
09308023
负责人：
INAGAKI Fuyuhiko
金额：
$ 21.5万
依托单位：
HOKKAIDO UNIVERSITY (1999)Tokyo Metropolitan Organization for Medical Research (1997-1998)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

We applied domain engineering to Ash/Grb2. We swapped nSH3 and cSH3 domains in Grb2 (SH3-SH2-SH3) and analyzed binding proteins using cell lysate from bovine brain. The binding proteins were similar to those for wild type Grb2, showing that each SH3 domain of Grb2 is independent . We also studied the phagocyte oxidation system in neutrophil. Phagocyte oxidation system consists of membrane bound gp91ィイD1phoxィエD1, p22phox and intracellular factor of p48ィイD1phoxィエD1, p67phox and p40ィイD1phoxィエD1. These intracellular factors have domain structure, comprising of SH3, PB1, PB2, TPR and PC motifs. We studied three dimensional structure of TRP motif in p67phox. Single crystal is now obtained and data collection has jut started using synchoroton radiationfacility.Phagocyte oxidation. System is triggered by binding of Rac(GTP) to TPR motif and p47ィイD1phoxィエD1 SH3 SH3 domain to p22ィイD1phoxィエD1 PRR. We designed novel signaling protein which consists of TPR motif directly connected to the SH3 domain of p47phox. The activity of this protein was studied in cell-free system containing membrane fraction of neutrophil and Rac-GTP form. The designed protein is quite active to generate super oxide. Thus, we have succeeded in preparing a protein based on the result of structural biology. We are going to extend this technique to prepare novel signaling proteins and regulate signal transduction system using the designed proteins.

我们将领域工程应用于Ash/Grb 2。我们交换了Grb 2中的nSH 3和cSH 3结构域（SH 3-SH 2-SH 3），并使用来自牛脑的细胞裂解物分析了结合蛋白。结合蛋白与野生型Grb 2相似，表明Grb 2的每个SH 3结构域是独立的。我们还研究了中性粒细胞吞噬细胞氧化系统。吞噬细胞氧化系统由膜结合的gp 91、p22 phox和胞内因子p48、p67 phox和p40等组成。这些胞内因子具有结构域结构，包括SH 3、PB 1、PB 2、TPR和PC基序。我们研究了p67 phox中TRP基序的三维结构。现在已经获得了单晶体，数据收集也刚刚开始使用同步辐射设备。系统由Rac（GTP）与TPR基序和p47 β D1 phox β D1 SH 3 SH 3结构域与p22 β D1 phox β D1 PRR的结合触发。我们设计了一种新的信号蛋白，它由TPR基序直接连接到p47 phox的SH 3结构域。在含有中性粒细胞膜组分和Rac-GTP形式的无细胞系统中研究了该蛋白的活性。所设计的蛋白质是相当活跃的产生超氧化物。因此，我们成功地制备了基于结构生物学结果的蛋白质。我们将扩展这项技术来制备新型信号蛋白并使用设计的蛋白质调节信号转导系统。