IDENTIFICATION AND CHARACTERIZATION OF Syk FAMIRY PROTEIN TYROSINE KINASE

Syk 家族蛋白酪氨酸激酶的鉴定和表征

基本信息

批准号：
09470043
负责人：
YAMAMURA Hirohei
金额：
$ 9.66万
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：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470043/
关键词：
Protein-tyrosine Syk vascular endothelial cells smooth muscle cells neurons tyrosine phosphorylated non-hematopoietic cells 血管平滑筋細胞 ZAP-70 神経成長円錐

项目摘要

Protein-tyrosine kinase Syk has been implicated in a variety of hematopoietic cell responses including immunoreceptor and integrin signaling. However, so far, there is no evidence of the expression and function of Syk in the non-hematopoietic tissues. in this study, we have purified 72kDa cytoplasmic protein-tyrosine kinase, having cross-reactivity with anti-Syk antibody, as a major cytosolic protein-tyrosine kinase from rat brain and blood cells-depleted rat liver. Partial sequence analysis of purified p72 kDa protein-derived peptides revealed that this protein is identical with Syk protein-tyrosine kinase. Immunohistochemical and RT-PCR northern analysis further demonstrated that Syk is expressed widely distributed in non-hematopoietic cells including vascular endothelial cells, smooth muscle cells, fibroblasts and neurons. Furthermore, Syk was significantly activated and tyrosine phosphorylated in response to a variety of ligands in non-hematopoietic cells. Thus, our results suggest the ubiquitous role of Syk in the signaling transduction of non-hematopoietic cells. We are currently attempting to establish several primary cultured cell lines derived from Syk-deficient mice. The genetic analysis of this mutant will provide a major tool in the physiological investigation of the functions of Syk. In addition to these results we studied about Syk from hematopoietic cells. The results obtained using hematopietic cells are quite useful to analyze the function of Syk in non-hematopoietic cells.

蛋白 - 酪氨酸激酶SYK已与包括免疫感受器和整联蛋白信号传导在内的多种造血细胞反应有关。但是，到目前为止，还没有证据表明SYK在非山摩托质组织中的表达和功能。在这项研究中，我们纯化了与抗SYK抗体的交叉反应性的72KDA细胞质蛋白 - 酪氨酸激酶，是大鼠脑和血细胞depledepled大鼠肝脏的主要胞质蛋白酪氨酸激酶。纯化的p72 kDa蛋白衍生的肽的部分序列分析表明，该蛋白与Syk蛋白 - 酪氨酸激酶相同。免疫组织化学和RT-PCR北部分析进一步表明，SYK在包括血管内皮细胞，平滑肌细胞，成纤维细胞和神经元在内的非山摩托细胞中广泛分布。此外，Syk被显着激活，酪氨酸响应于非造成杂物细胞的多种配体而磷酸化。因此，我们的结果表明，Syk在非杂造细胞的信号转导中无处不在。我们目前正在尝试建立从SYK缺陷小鼠衍生的几种原发性培养细胞系。该突变体的遗传分析将为SYK功能的生理研究提供主要工具。除这些结果外，我们还研究了来自造血细胞的SYK。使用造血细胞获得的结果对于分析SYK在非脊髓性细胞中的功能非常有用。