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

Study on the buildup principle of the plasma membrane signaling domains.

质膜信号结构域构建原理的研究。

基本信息

批准号：
11680658
负责人：
SATO Satoshi
金额：
$ 2.18万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

The plasma membrane contains various forms of invaginations with apparently different physical building principles. To specify the mechanisms of domain formation, particularly those for clathrin-coated pits and caveolae, the dynamical characterization of the lattice assembly should be an important approach. We addressed the effects of poly (ethylene glycol) cholesteryl ether (PEG-Chol) on the structure/function of clathrin-coated pit and caveolae. Addition of the compound to cultured cells induced progressive smoothening of the surface. In these cells were accumulated opened clathrin-coated pits, which ultimately became flat on occasion of the cell rupture by slightly more PEG-Chol. These results strongly suggested that coated-pit elastically differentiates from the bulk membrane phase and also from caveolae. Next, we analyzed the effect of accumulated membrane stress. Whereas simple incubation of PEG-Chol gave no morphological effect, we found numerous plasma membrane tubules decorate … More d with biotinylated PEG-Chol (bPEG-Chol) and SA were introduced. The tubules were homogeneous in diameter (d【approximately equal】100 nm) and contained actin filaments. Biochemically, bPEG-Chol induced tyrosinephosphorylation of proteins. The analysis suggested that local accumulation of lipid-packing defects evoked signaling reaction (s) that mobilize actin to reduce the stress from the main cell body. Next, we describe the responses of K562 human leukemia cells that were treated with okadaic acid, an inhibitor of type 1 and 2A protein dephosphatases, to physical stress on the plasma membrane. While OKA alone induced patchy accumulation of cortical F-actin, application of mechanical stresses induced budding of membrane-bound body containing the whole cell F-actin. The formation of this actin-rich body was prevented by modification of F-actin and also by protein kinase-inhibitors. Moreover, when a non-receptor tyrosine kinase Fyn was knocked-out, such change did not occur. In this project, we suggest the presence of the specific mechanical linkage between actin microfilaments and a defined set of membrane organelles or surfaces proteins in OKA-treated cells. Less

质膜包含各种形式的内陷，具有明显不同的物理构建原理。要指定的机制域的形成，特别是那些网格蛋白涂层的坑和小窝，动态表征的晶格组装应该是一个重要的方法。我们解决了聚（乙二醇）胆固醇醚（PEG-Chol）的结构/功能的网格蛋白包被的小窝和小窝的影响。向培养的细胞中添加化合物诱导表面进行性平滑。在这些细胞中积累了开放的网格蛋白包被的凹坑，当细胞破裂时，这些凹坑最终变得平坦。这些结果强烈表明，包被坑弹性区分从散装膜相，也从小窝。接下来，我们分析了累积膜应力的影响。而PEG-Chol的简单孵育没有形态学影响，我们发现许多质膜小管装饰 ...更多信息 d与生物素化的聚乙二醇胆固醇（bPEG-Chol）和SA的反应。小管直径均匀（d[约等于]100 nm），含有肌动蛋白丝。生物化学上，bPEG-Chol诱导蛋白质的酪氨酸磷酸化。分析表明，局部积累的脂质包装缺陷诱发信号反应，动员肌动蛋白，以减少来自主细胞体的压力。接下来，我们描述了K562人白血病细胞的反应，用冈田酸，1型和2A型蛋白脱磷酸酶的抑制剂，对质膜的物理应力处理。虽然OKA单独诱导皮质F-肌动蛋白的斑片状积累，机械应力的应用诱导出芽的膜结合体包含整个细胞F-肌动蛋白。这种肌动蛋白丰富的身体的形成被阻止修改的F-肌动蛋白和蛋白激酶抑制剂。此外，当敲除非受体酪氨酸激酶Fyn时，不发生这种变化。在这个项目中，我们建议在OKA处理的细胞中肌动蛋白微丝和一组确定的膜细胞器或表面蛋白之间存在特定的机械连接。少