development of preparation of modified tRNA using protein

利用蛋白质制备修饰tRNA的进展

• 批准号: 24655145
• 负责人: TANAKA Yoshikazu
• 金额: $ 2.58万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24655145/
• 关键词: tRNA; 精製; 修飾; 結晶化; 結晶構造解析; 結晶

In this study, I tried to establish conventional method to prepare highly pure modified tRNA from E. coli cell using tRNA-protein complex spontaneously formed in the cells. First, I established preparation system for tRNA(Met) using E. coli tRNA(Met) acetyltransferase. For this purpose, I selected K205A mutant, which has high affinity for tRNA. Moreover, crystals of the purified tRNA(Met) were obtained. The crystals diffracted about 6-angstrom resolution at synchrotron radiation facility. Furthermore, co-expression system of selenium introducing enzyme with tRNA(Gln), tRNA(Glu), and tRNA(Lys) in E. coli were constructed, and pure tRNAs were prepared from the tRNA-protein complex spontaneously formed in the E. coli cell.

本研究试图建立一种从大肠杆菌中制备高纯度修饰tRNA的常规方法。大肠杆菌细胞使用细胞内自发形成的tRNA-蛋白质复合物。首先，建立了大肠杆菌tRNA（Met）的制备体系; coli tRNA（Met）乙酰转移酶。为此，我选择了K205 A突变体，它对tRNA具有高亲和力。此外，获得纯化的tRNA（Met）的晶体。该晶体在同步辐射装置上衍射的分辨率约为6埃。此外，还建立了硒导入酶与tRNA（Gln）、tRNA（Glu）、tRNA（Lys）在大肠杆菌中的共表达系统。coli中自发形成的tRNA-蛋白质复合物制备纯的tRNA。coli细胞。

项目成果

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Structural and genomic DNA analysis of a putative transcription factor SCO5550 from Streptomyces coelicolor A3 (2) : Regulating the expression of gene sco5551 as a transcriptional activator with a novel diner shape

天蓝色链霉菌 A3 的假定转录因子 SCO5550 的结构和基因组 DNA 分析 (2)：调节 sco5551 基因的表达，作为具有新型晚餐形状的转录激活剂

• DOI: 10.1016/j.bbrc.2013.04.017
• 发表时间: 2013
• 期刊: Biochemical and Biophysical Research Communications
• 影响因子: 3.1
• 作者: Takeshi Hayashi;Yoshikazu Tanaka;Naoki Sakai;Nobuhisa Watanabe;Tomohiro Tamura;Isao Tanaka;Min Yao
• 通讯作者: Min Yao

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

Application of Photoactive Yellow Protein as a Photoresponsive Module for Controlling Hemolytic Activity of Staphylococcal α-Hemolysin

光活性黄色蛋白作为光响应模块在控制葡萄球菌α-溶血素溶血活性中的应用

• DOI: 10.1039/c2cc18118e
• 发表时间: 2012
• 期刊: Chemical Communications
• 影响因子: 4.9
• 作者: M. Ui;et al.
• 通讯作者: et al.

Crystal structure analysis of a DING protein, International life-science symposium for young scientists

DING 蛋白的晶体结构分析，国际青年科学家生命科学研讨会

• 发表时间: 2014
• 作者: Zuoqi Gai;Akiyoshi Nakamura;Yoshikazu Tanaka;Nagisa Hirano;Isao Tanaka;and Min Yao
• 通讯作者: and Min Yao