Role of cytoskeletal proteins phosphorylation on their cellular functions.

细胞骨架蛋白磷酸化对其细胞功能的作用。

基本信息

批准号：
04454177
负责人：
INAGAKI Masaki
金额：
$ 4.22万
依托单位：
Tokyo Metropolitan Institute of Gerontology
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

Phosphorylation and dephosphorylation of proteins dynamically alter the structure and function of proteins and are regarded as a form of an on/off switch regulating various vital phenomena. If the phosphorylation and dephosphorylation of proteins could be visualized, especially in terms of the distribution, site and transition of each reaction, the relevant and various vital phenomena, that currently can be investigated merily by indirect means, might be more revealing. Based on this concept, we prepared a group of antibodies that can identify whether a cytoskeletal protein is phosphorylated or dephosphorylated.We developed four antibodies which distinguish phosphorylated states of Thr7, Ser8, Ser13 and Ser34 in glial fibrillary acidic protein (GFAP), a protein constituted of glial filaments of astroglial cells. Immunofluorescence microscopy shows that Ser8 residues in all cytoplasmic glial filaments are initially phosphorylated when the astroglial cells enter mitosis. In cytokinesis, the phospho-Ser8 residues become dephosphorylated, whereas Thr7, Ser13 and Ser34 in the filaments at the cleavage furrow become the preferred sites of phosphorylation.Such being the case, at least two different types of protein kinases act as mitotic glial filament kinases, at a different time schedule during mitosis. A phosphatase (s) acts as a mitotic glial filament phosphatase, at the time of meta-anaphase transition.

蛋白质的磷酸化和去磷酸化动态改变了蛋白质的结构和功能，被视为调节各种重要现象的开/关开关的一种形式。如果可以将蛋白质的磷酸化和去磷酸化可视化，尤其是在每个反应的分布，位点和过渡方面，相关和各种重要现象，目前可以通过间接手段进行合并的相关和各种重要现象，可能会更揭示。基于这个概念，我们准备了一组抗体，这些抗体可以识别细胞骨架蛋白是磷酸化还是去磷酸化。我们开发了四种抗体，这些抗体在胶质纤维纤维蛋白（GFAP）中区分Thr7，Ser8，Ser8，Ser13和Ser34的磷酸化态，，一种由星形胶质细胞的神经胶质构成。免疫荧光显微镜表明，当星形胶质细胞进入有丝分裂时，所有细胞质神经胶质细丝中的Ser8残基最初磷酸化。在细胞因子中，磷酸-SER8残基被脱磷酸化，而在裂解沟的细丝中，Thr7，Ser13和Ser34成为磷酸化的首选位点。与之相比，至少是两种不同类型的蛋白质激酶起作用。，在有丝分裂期间的不同时间表。磷酸酶在荟萃递变时充当有丝分裂神经胶质丝磷酸酶。