Identification and characterization of novel kinase, AKB14-3-3-1, which is a possible target of sleeping sickness.

新型激酶 AKB14-3-3-1 的鉴定和表征，该激酶是昏睡病的可能靶标。

• 批准号: 23590500
• 负责人: INOUE Masahiro
• 金额: $ 3.33万
• 依托单位: Kurume University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23590500/
• 关键词: Tb14-3-3; 蛋白リン酸化酵素; 細胞分裂; 細胞骨格; リン酸化モチーフ; kinase substrate Ab; cytokinesis; inhibitor; complex; AKB14-3-3-1; Trypanosoma brucei; 14-3-3; kinase; cell division; 2014/3/3; cell divison

Proteomics approach of searching Tb14-3-3 binding proteins identified a novel associated kinase of Tb14-3-3 (AKB1). Tb14-3-3 directly and specifically binds to AKB1 utilizing an amphipathic groove structure of Tb14-3-3. Critical amino acid motif forAKB1 substrate recognition was identified and in vivo AKB1 kinase activity was successfully monitored with phosphorylated substrate-specific antibody. Majority of AKB1 is associated with cytoskeleton and most of AKB1 substrates are also identified in the detergent-insoluble fraction of cell lysate. AKB1 knockdown or overexpression of wild type AKB1 but not kinase-dead AKB1, deregulated cytokinesis and cell division, suggesting that kinase activity of AKB1 is crucial for growth. The results from this study suggest that AKB1 associated with Tb14-3-3 modulates cytokinesis and cell cycle progression in Trypanosoma brucei by phosphorylating cytoskeleton-associated proteins.

通过蛋白质组学方法寻找Tb 14 -3-3结合蛋白，发现了一个新的Tb 14 -3-3相关激酶（AKB 1）。Tb 14 -3-3利用Tb 14 -3-3的两亲沟结构直接特异性结合AKB 1。确定了AKB 1底物识别的关键氨基酸基序，并成功地用磷酸化底物特异性抗体监测了体内AKB 1激酶活性。大多数AKB 1与细胞骨架相关，并且大多数AKB 1底物也在细胞裂解物的洗涤剂不溶性部分中鉴定。野生型AKB 1的AKB 1敲除或过表达（但非激酶死亡的AKB 1）会解除胞质分裂和细胞分裂的调节，这表明AKB 1的激酶活性对生长至关重要。这项研究的结果表明，与Tb 14 -3-3相关的AKB 1通过磷酸化胞浆素相关蛋白来调节布氏锥虫的胞质分裂和细胞周期进程。

项目成果

AKB1(associated kinase of Tb14-3-3-1)のin vitro酵素活性

AKB1（Tb14-3-3-1 相关激酶）的体外酶活性

• 发表时间: 2014
• 作者: 井上雅広;安田幸一;上村春樹
• 通讯作者: 上村春樹

14-3-3タンパク質によるDAPK2活性制御機構の解析

14-3-3蛋白对DAPK2活性调控机制分析

• 发表时间: 2013
• 作者: 太田玲奈;松田真弥;井上雅広;辻明彦;湯浅恵造
• 通讯作者: 湯浅恵造

Associated kinase of Trypansoma brucei 14-3-3(AKB1)の分子生物学

布氏锥虫相关激酶14-3-3(AKB1)的分子生物学

• 发表时间: 2012
• 作者: Tanabe K;Zollnerc G;Vaughanc JA;Sattabongkotc J;Khuntiratc B;Honma H;Mita T;Tsuboi T;Coleman R;井上雅広
• 通讯作者: 井上雅広

AKB14-3-3 (Associated kinase of Trypanosoma brucei 14-3-3)

AKB14-3-3（布氏锥虫 14-3-3 相关激酶）

• 作者: 葛西真治;駒形修;糸川健太郎;小林睦生;冨田隆史;井上雅広
• 通讯作者: 井上雅広

Associated kinase of Trypanosoma brucei 14-3-3-1(AKB1)の分子生物学

布氏锥虫相关激酶14-3-3-1(AKB1)的分子生物学

• 发表时间: 2012
• 作者: 平田晃一;駒形修;糸川健太郎;山本敦司;冨田隆史;葛西真治;新倉 保，井上信一，小林富美惠;井上雅広
• 通讯作者: 井上雅広