The study on the multi-molecular reaction mechanism of transamidosome for Gln-tRNA(Gln) synthesis

转酰胺体合成Gln-tRNA(Gln)的多分子反应机制研究

• 批准号: 21370041
• 负责人: YAO Min
• 金额: $ 12.23万
• 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: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21370041/
• 关键词: X 線結晶解析; 多段階反応の協調; トランスアミドソーム; GatCAB; tRNA; GluRS; AspRS; 多分子協調機構; X線結晶解析

In order to elucidate the multi-molecular reaction mechanism of transamidosome in the indirect synthetic pathway of aminoacyl tRNA (Gln-tRNA^Gln, Asn-tRNA^Asn), we over-expressed, purified 6 components of 2 transamidosomes from S. aureus and P. aeruginosa, respectively, and formed 2 transamidosomes in vitro. We succeeded in the structural analysis of Asn-tRNA^Asn transamidosomes of P.aeruginosa. The structure of Asn-tRNA^Asn transamidosomes consisted of tRNA^Asn, ND-AspRS and GatCAB with a rate of 2:2:2, according with the result of the biochemistry experiment. The structure shown that acceptor stem of tRNA^Asn was located in the active site of ND-AspRS, representing a tRNA-binding situation before aminoacylation (mischarge) of the tRNA^Asn. After the aminoacylation, the acceptor stem of tRNAAsn flipped toward the active site of GatCAB for transamidation.

为了阐明转酰胺体在氨酰tRNA（Gln-tRNA^Gln，Asn-tRNA^Asn）间接合成途径中的多分子反应机理，我们从S.金黄色葡萄球菌和铜绿假单胞菌，并形成2个transamidosomes在体外。我们成功地对铜绿假单胞菌的Asn-tRNA、Asn转酰胺体进行了结构分析。Asn-tRNA、Asn反式酰胺体的结构由tRNA、Asn、ND-AspRS和GatCAB组成，比例为2：2：2，与生化实验结果一致。结构表明，tRNA^Asn的受体茎位于ND-AspRS的活性位点，代表tRNA^Asn氨酰化（错电荷）之前的tRNA结合情况。在氨酰化后，tRNAAsn的受体茎翻转到GatCAB的活性位点进行转酰胺。

项目成果

The binding mechanism of eIF2β with its partner proteins, eIF5 and eIF2Bε, Biochem

eIF2β与其伴侣蛋白eIF5和eIF2Bε的结合机制，Biochem

• DOI: 10.1016/j.bbrc.2012.05.155
• 发表时间: 2012
• 期刊: Biophys. Res. Commun
• 作者: Gai Z;Kitagawa Y;Tanaka Y;Shimizu N;Komoda K;Tanaka I;Yao M
• 通讯作者: Yao M

The Recognition Mechanism of eIF2β for its partner proteins eIF5 and eIF2Bε

eIF2β对其伴侣蛋白eIF5和eIF2Bε的识别机制

• 发表时间: 2011
• 作者: 神川龍馬;稲垣祐司;Zuoqi Gai
• 通讯作者: Zuoqi Gai

Snapshots of dynamics in synthesizing N(6)-isopentenyladenosine at the tRNA anticodon.

• DOI: 10.1021/bi900337d
• 发表时间: 2009-06-16
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Chimnaronk, Sarin;Forouhar, Farhad;Sakai, Junichi;Yao, Min;Tron, Cecile M.;Atta, Mohamed;Fontecave, Marc;Hunt, John F.;Tanaka, Isao
• 通讯作者: Tanaka, Isao

RNA helicase module in an acetyltransferase that modifies a specific tRNA anticodon

• DOI: 10.1038/emboj.2009.69
• 发表时间: 2009-05-06
• 期刊: EMBO JOURNAL
• 影响因子: 11.4
• 作者: Chimnaronk, Sarin;Suzuki, Tateki;Tanaka, Isao
• 通讯作者: Tanaka, Isao

Crystal Structures of the Multidrug Binding Repressor Corynebacterium glutamicum CgmR in Complex with Inducers and with an Operator

• DOI: 10.1016/j.jmb.2010.07.042
• 发表时间: 2010-10-22
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Itou, Hiroshi;Watanabe, Nobuhisa;Tanaka, Isao
• 通讯作者: Tanaka, Isao