Analysis of translation mechanism for selenoprotein synthesis

硒蛋白合成翻译机制分析

• 批准号: 22770093
• 负责人: OSE Toyoyuki
• 金额: $ 2.75万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22770093/
• 关键词: 翻訳マシーナリ; RNA調製; 蛋白質調製; 翻訳マシーン; リボソーム

We successfully prepared protein and RNA molecules which are required for selecocystein incorporation on ribosome. These molecules are SBP2, L30, EFSec, SEICIS-RNA, tRNASec and subjected to biochemical and biophysical experiments. Crystal structure of L30 could be analyzed and published. Since we originally succeeded generating functional tRNASec, binding assay among tRNASec, EFSec, and SBP2 in the presence of specific amino acid are now in progress. We observed the significant binding affinity(K_d=229 nM) between the wild type and GDP using ITC, the way to prepare the molecule was thus confirmed. Furthermore, mutants generated for the purpose of crystallization were proved that they have proper secondary structures

我们成功地制备了选择半胱氨酸与核糖体结合所需的蛋白质和RNA分子。这些分子分别是SBP2、L30、EFSec、SEICIS-RNA、tRNASec，并进行了生化和生物物理实验。L30的晶体结构可以分析并发表。由于我们最初成功地生成了功能性tRNASec，目前正在进行tRNASec、EFSec和SBP2在特定氨基酸存在下的结合实验。我们利用ITC观察到野生型与GDP具有显著的结合亲和力（K_d=229 nM），从而确定了该分子的制备方法。此外，为结晶目的而产生的突变体被证明具有适当的二级结构

项目成果

10. Mechanistic analysis of the 21st amino acid incorporation system in eukaryotes

10.真核生物第21种氨基酸掺入系统的机理分析

• 发表时间: 2012
• 作者: Akiko Kawaguchi;Toyoyuki Ose;Min Yao;Isao Tanaka
• 通讯作者: Isao Tanaka

Structure analysis of SelB in complex with SECIS mRNA from Moorella thermoacetica

SelB 与热乙酸穆尔氏菌 SECIS mRNA 复合物的结构分析

• 发表时间: 2010
• 作者: Toyoyuki Ose;et.al.
• 通讯作者: et.al.