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Studies on the function of a centrosome and Golgi localized giant anchoring protein CG-NAP

中心体和高尔基体定位巨型锚定蛋白CG-NAP的功能研究

基本信息

批准号：
13480209
负责人：
ONO Yoshitaka
金额：
$ 3.2万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13480209/
关键词：
protein kinases PKN PKC anchoring proteins CG-NAP centrosome Golgi apparatus

项目摘要

Protein kinases are the key components in the intracellular signal transduction. The signaling specificity of the protein kinases may be mediated by the targeting of them to the specific micro-organelles through the anchoring proteins. CG-NAP (centrosome and Golgi localized PKN-associated protein) is a coiled-coil protein identified as a binding protein for the regulatory domain of PKN that has a catalytic domain highly homologous to PKC in the carboxyl-terminal region and a unique regulatory domain in the amino-terminal region. In this study, we have analyzed the function of CG-NAP as an anchoring protein and obtained the results as follows.1. CG-NAP is a 450-kDa coiled-coil protein and is localized to centrosome throughout the cell cycle, the midbody at telophase, and the Golgi apparatus at interphase.2. In addition to PKN, CG-NAP associates with protein kinase A, PKCε, and protein phosphatases (PP1 and PP2A).3. CG-NAP anchors non/hypo-phosphrylated PKCε at Golgi/centrosome area during maturation, and serves as a scaffold for the phosphorylation reaction.4. CG-NAP provides sites for microtubule nucleation in the mammalian centrosome by anchoring γ-tubulin ring complex.5. The carboxyl-terminal region of CG-NAP is necessary for the localization to centrosome.

蛋白激酶是细胞内信号转导的关键组分。蛋白激酶的信号传导特异性可以通过锚定蛋白将它们靶向特定的微细胞器来介导。CG-NAP（centrosome and Golgi localized PKN-associated protein）是一种卷曲螺旋蛋白，被鉴定为PKN调节结构域的结合蛋白，其羧基端区域具有与PKC高度同源的催化结构域，氨基端区域具有独特的调节结构域。本研究对CG-NAP作为锚定蛋白的功能进行了分析，得到以下结果. CG-NAP是一个分子量为450-kDa的卷曲螺旋蛋白，在细胞周期中定位于中心体，在细胞分裂末期定位于中间体，在分裂间期定位于高尔基体.除PKN外，CG-NAP还与蛋白激酶A、PKCε和蛋白磷酸酶（PP 1和PP 2A）结合. CG-NAP在成熟过程中将非磷酸化/低磷酸化的PKCε锚定在高尔基体/中心体区域，并作为磷酸化反应的支架. CG-NAP通过锚定γ-微管蛋白环复合物为哺乳动物中心体中的微管成核提供位点. CG-NAP的羧基末端区域对于定位于中心体是必需的。