Comprehensive analysis of molecules related for antigen receptor signaling

抗原受体信号传导相关分子综合分析

• 批准号: 16300093
• 负责人: HATAKEYAMA Shigetsugu
• 金额: $ 9.66万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16300093/
• 关键词: proteome; ubiquitylation; phosphorylation; lymphocyte; mass spectrometry

This application is to analyze comprehensively about post-translational modification regarding to phosphorylation and ubiquitylation using affinity-chromatography. The system we analyzed is signal transduction system of antigen receptor signals in immune cells, especially phosphorylation and ubiquitylation. The purpose of this project is to clarify the down-stream signal transduction molecules comprehensively from the point of protein modifications.Ubiquitylation and phosphorylation are implicated in the many intracellular phenomenons including signal transduction such as TCR,BCR,FcR signals. We analyze what molecules are ubiquitylated by collecting ubiquitylated proteins with anti-ubiquitin affinity chromatography. We tried to concentrate ubiquitylated proteins and ubiquitin-interacting proteins, to separate by SDS-PAGE, and to analyze these proteins by mass spectrometer. This analysis showed about 700 ubiquitin-related proteins. Furthermore we tried to concentrate tyrosine-phosphorylated proteins by anti-phosphotyrosine and to analyze these proteins by mass spectrometer. This analysis showed about 200 proteins (including novel proteins) which are related to tyrosine phosphorylation. It is significant to clarify the etiology of diseases such as autoimmune diseases and allergic diseases by connecting between protein modifications (such as phosphorylation and ubiquitylation) and activating signals by antigen receptors.

本申请旨在利用亲和色谱法全面分析磷酸化和泛素化的翻译后修饰。我们分析的系统是免疫细胞中抗原受体信号的信号转导系统，特别是磷酸化和泛素化。本项目的目的是从蛋白质修饰的角度全面阐明下游信号转导分子。泛素化和磷酸化与TCR、BCR、FcR信号等信号转导等多种细胞内现象有关。我们通过使用抗泛素亲和层析收集泛素化蛋白质来分析哪些分子被泛素化。我们尝试浓缩泛素化蛋白和泛素相互作用蛋白，通过SDS-PAGE分离，并通过质谱仪分析这些蛋白。该分析显示大约 700 种泛素相关蛋白。此外，我们尝试通过抗磷酸酪氨酸浓缩酪氨酸磷酸化蛋白质，并通过质谱仪分析这些蛋白质。该分析显示了大约 200 种与酪氨酸磷酸化相关的蛋白质（包括新蛋白质）。通过蛋白质修饰（如磷酸化和泛素化）与抗原受体激活信号之间的联系，对于阐明自身免疫性疾病和过敏性疾病等疾病的病因具有重要意义。

项目成果

AIRE functions as an E3 ubiquitin ligase.

• DOI: 10.1084/jem.20031291
• 发表时间: 2004-01-19
• 期刊: The Journal of experimental medicine
• 作者: Uchida D;Hatakeyama S;Matsushima A;Han H;Ishido S;Hotta H;Kudoh J;Shimizu N;Doucas V;Nakayama KI;Kuroda N;Matsumoto M
• 通讯作者: Matsumoto M

CHIP promotes proteasomal degradation of familial ALS-linked mutant SOD1 by ubiquitinating Hsp/Hsc70

• DOI: 10.1111/j.1471-4159.2004.02486.x
• 发表时间: 2004-07-01
• 期刊: JOURNAL OF NEUROCHEMISTRY
• 影响因子: 4.7
• 作者: Urushitani, M;Kurisu, J;Takahashi, R
• 通讯作者: Takahashi, R

Ubiquitylation and degradation of polo-like kinase SNK by hVPS18, a RING-H2 type ubiquitin ligase.

hVPS18（一种 RING-H2 型泛素连接酶）对 polo 样激酶 SNK 进行泛素化和降解。

• 发表时间: 2005
• 期刊: J.Biol.Chem. 280
• 作者: Yogosawa;S.
• 通讯作者: S.

Large-scale analysis of the human ubiquitin-related proteome

• DOI: 10.1002/pmic.200401280
• 发表时间: 2005-11-01
• 期刊: PROTEOMICS
• 影响因子: 3.4
• 作者: Matsumoto, M;Hatakeyama, S;Nakayama, KI
• 通讯作者: Nakayama, KI

Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27Kip1 at G1 phase

• DOI: 10.1038/ncb1194
• 发表时间: 2004-12-01
• 期刊: NATURE CELL BIOLOGY
• 影响因子: 21.3
• 作者: Kamura, T;Hara, T;Nakayama, KI
• 通讯作者: Nakayama, KI