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Analysis of the Rap2- MAP4K signaling system

Rap2-MAP4K信号系统分析

基本信息

批准号：
16590251
负责人：
KARIYA Ken-ichi
金额：
$ 1.73万
依托单位：
University of the Ryukyus
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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/en/grant/KAKENHI-PROJECT-16590251/
关键词：
signal transduction Rap2 HGK TNIK MINK MAP4K JNK Rasファミリー

项目摘要

We have recently found that a Ras-like small GTP-binding protein Rap2 interacts specificaly with three protein kinases : (1)Nck-interacting kinase (NIK), also known as HPK/GCK-like kinase (HGK) or mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4) ; (2)Traf-and Nck-interacting kinase (TNIK) ; and (3)Misshapen/NIKs related kinase (MINK). These kinases belong to the GCK family of the STE20 group kinases. The STE20 group kinases share kinase domains homologous to that of STE20, a MAP4K in budding yeast. STE20 group kinases in mammals regulate stress-activated mitogen-activated protein kinases (MAPKs) such as c-Jun N-terminal kinase (JNK) and p38MAPK. NIK/HGK/MAP4K4, TNIK, and MINK share similar N-terminal kinase domains and C-terminal regulatory domains. Rap2 binds to the C-terminal regulatory domains of these kinases in a GTP-dependent manner. The C-terminal regulatory domain is also termed the CNH domain (citron homology domain), since similar structure is found in citron, an effector molecule of Rho family small GTP-binding proteins such as Cdc42 and Rac. NIK/HGK/MAP4K4, TNIK, and MINK did not interact with Ras or Rap1. Thus, we believe that NIK/HGK/MAP4K4, TNIK, and MINK are specific effector molecules of Rap2 and are components of a new signaling system, which we term the Rap2-MAP4K system. To identify molecules that are upstream or downstream of this Rap2-MAP4K system, we utilized GST-TNIK affinity chromatography coupled with mass spectrometry, and identified a scaffold protein with tricopeptide repeats, ankyrin repeats, a coiled-coil region and a PDZ-binding motif. The analysis also identified TNIK itself as a TNIK-binding protein, suggesting that the system contains dimmer or oligomer of MAP4K. We also utilized the differential two-dimensional gel electrophoresis and found potential phosphoproteins regulated by TNIK.

我们最近发现Ras样小GTP结合蛋白Rap 2与三种蛋白激酶特异性相互作用：（1）Nck相互作用激酶（NIK），也称为HPK/GCK样激酶（HGK）或促分裂原活化蛋白激酶4（MAP 4K 4）;（2）Traf和Nck相互作用激酶（TNIK）;和（3）Misshapen/NIK相关激酶（MINK）。这些激酶属于STE 20组激酶的GCK家族。STE 20组激酶共享与STE 20（芽殖酵母中的MAP 4K）同源的激酶结构域。哺乳动物中的STE 20组激酶调节应激激活的有丝分裂原激活蛋白激酶（MAPKs），例如c-Jun N末端激酶（JNK）和p38 MAPK。NIK/HGK/MAP 4K 4、TNIK和MINK共享相似的N-末端激酶结构域和C-末端调节结构域。Rap 2以GTP依赖性方式结合这些激酶的C-末端调节结构域。C-末端调节结构域也被称为CNH结构域（香橼同源结构域），因为在香橼中发现了类似的结构，香橼是Rho家族小GTP结合蛋白如Cdc 42和Rac的效应分子。NIK/HGK/MAP 4K 4、TNIK和MINK不与Ras或Rap 1相互作用。因此，我们认为NIK/HGK/MAP 4K 4、TNIK和MINK是Rap 2的特异性效应分子，并且是一种新的信号系统的组分，我们称之为Rap 2-MAP 4K系统。为了鉴定Rap 2-MAP 4K系统的上游或下游分子，我们利用GST-TNIK亲和层析结合质谱，并鉴定了具有三肽重复序列、锚蛋白重复序列、卷曲螺旋区域和PDZ结合基序的支架蛋白。该分析还将TNIK本身鉴定为TNIK结合蛋白，这表明该系统包含MAP 4K的二聚体或寡聚体。我们还利用差异双向凝胶电泳，发现潜在的磷蛋白TNIK调节。