Molecular basis of the adaptor domains from AAA-ATPases for locating organelles.

用于定位细胞器的 AAA-ATP 酶接头结构域的分子基础。

• 批准号: 16570136
• 负责人: HIROAKI Hidekazu
• 金额: $ 2.3万
• 依托单位: Yokohama City University
• 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-16570136/
• 关键词: functional protein domains; Nuclear magnetic resonance (NMR); protein domain anatomy; late endosome; tubulin; cell skeleton; MIT domain; sequence identity; プロテアソーム; ポリユビキチン; ペルオキシソーム; X線結晶構造解析

AAA-ATPases are well conserved molecular machines from bacteria to human, which alters the conformations and/or multimerization states of substrate proteins with an ATP-depending manner. AAA-ATPases are characterized as harboring one or several "AAA" domains at their central region of the molecules, whereas various N-terminus domains may be responsible for differentiating their specific functions. I have hypothesized that some of them can be responsible not only for their substrate specificity, but also for their subcellular location. In order to solve this problem, so-called "protein domain anatomy" approach was taken.As a result, first we have identified a novel domain from the PEX1 N-terminal region. Surprisingly, although below 15% of sequence identity found to VCP and NSF, the structure of PEX1NTD is very similar to those corresponding domains. Second, we have identified the solution structures from N-terminal MIT domain of Vps4 type I AAA-ATPase and its distant homolog katanin p60. These domains adopted into an up-and-down three helix bundle fold with a characteristic cleft on the molecular surface, which may act as a ligand binding site, katanin p60 N-terminal domain was associated with a coiled-coil region, which should be designed prior to structural analysis. Some other AAA-ATPases require a shuttle factor molecule instead of its own adaptor domain. From one of them, we determined the solution structure of UBA domain from Dsk2 protein, an adaptor of 19S proteosome, in complex with ubiquitin. In addition, molecular rigidity and flexibility of K48- and K63-linked tetra ubiquitin chain were compared by using solution NMR.Finally, two domains of the five examined systems were found to have affinities to phosphoinositides. This suggested the domains share a function for associating organellar membranes.

从细菌到人类，aaa - atp酶都是保守的分子机器，它以atp依赖的方式改变底物蛋白的构象和/或聚合状态。AAA- atp酶的特点是在分子的中心区域含有一个或几个“AAA”结构域，而不同的n端结构域可能负责区分它们的特定功能。我假设它们中的一些不仅对它们的底物特异性负责，而且对它们的亚细胞位置负责。为了解决这一问题，采用了所谓的“蛋白质结构域解剖学”方法。因此，我们首先从PEX1 n端区域确定了一个新的结构域。令人惊讶的是，尽管PEX1NTD与VCP和NSF的序列一致性低于15%，但其结构与相应的结构域非常相似。其次，我们确定了Vps4 I型aaa - atp酶的n端MIT结构域及其远端同系物katanin p60的溶液结构。这些结构域在分子表面呈上下三螺旋束折叠，并具有特征性的裂缝，可能作为配体结合位点，katanin p60 n端结构域与一个盘状盘状区域相关联，在进行结构分析之前应进行设计。其他一些aaa - atp酶需要穿梭因子分子而不是它自己的适配器结构域。从其中一份样品中，我们确定了19S蛋白体适配器Dsk2蛋白与泛素复合物中UBA结构域的溶液结构。此外，利用溶液核磁共振比较了K48-和k63 -连接的四泛素链的分子刚性和柔韧性。最后，五个检查系统的两个区域被发现有亲和力的磷酸肌苷。这表明这些结构域具有联合细胞器膜的功能。

项目成果

High-throughput construction method for expression vector of peptides for NMR study suited for isotopic labeling.

• DOI: 10.1093/protein/gzh044
• 发表时间: 2004-04
• 期刊: Protein engineering, design & selection : PEDS
• 作者: T. Tenno;Natsuko Goda;Y. Tateishi;H. Tochio;M. Mishima;H. Hayashi;M. Shirakawa;H. Hiroaki

ペルオキシソーム因子PEX1 ATPaseのN末端ドメインの立体構造が示したAAA ATPase間の進化的類縁関係

过氧化物酶体因子 PEX1 ATP 酶 N 端结构域的三维结构揭示了 AAA ATP 酶之间的进化关系

• 发表时间: 2006
• 期刊: 生物物理 46(3)
• 作者: 廣明 秀一;塩澤久美子;富井健太郎
• 通讯作者: 富井健太郎

Structural characterization of the MIT domain from human Vps4b

• DOI: 10.1016/j.bbrc.2005.06.110
• 发表时间: 2005-08-26
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Takasu, H;Jee, JG;Hiroaki, H
• 通讯作者: Hiroaki, H

Structure of the N-terminal domain of PEX1 AAA-ATPase : characterization of a putative adaptor-binding domain.

PEX1 AAA-ATPase N 端结构域的结构：假定的接头结合域的表征。

• 发表时间: 2004
• 期刊: Journal of Biological Chemistry 279
• 作者: Shiozawa;K.;Maita;N.;Tomii;K.;Seto;A.;Goda N.;Akiyama;Y.;Shimizu;T.;Shirakawa;M.;Hiroaki;H.
• 通讯作者: H.

ヒトVps4bのMITドメインと二価または三価の金属イオン複合体の構造的特徴およびヒトVps4bのMITドメインによるホスファチジルイノシトールリン酸認識機構

人Vps4b MIT结构域与二价或三价金属离子复合物的结构特征以及人Vps4b MIT结构域对磷酸磷脂酰肌醇的识别机制

• 发表时间: 2004