CRYSTAL STRUCTURE DETERMINATION OF QUEOSINE-SYNTHESIS AND OTHER TRNA MODIFICATIO

奎辛合成和其他 TRNA 修饰的晶体结构测定

• 批准号: 7722119
• 负责人: MANAL A SWAIRJO
• 金额: $ 0.02万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722119
• 关键词: Anabolism; Antibiotics; Anticodon; Automation; Biochemical; Codon Nucleotides; Complex; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Data; Drug Delivery Systems; Enzymes; Funding; Genomics; Grant; Institution; Ligands; Methods; Modification; Pathway interactions; Prokaryotic Cells; Research; Research Personnel; Resources; Ribosomes; Shigella Infections; Source; Structure; Technology; Time; Transfer RNA; Translations; United States National Institutes of Health; analog; beamline; comparative

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have used SSRL beam time on this proposal to conduct crystallographic studies on enzymes responsible for the biosynthesis of two tRNA modifications: queosine (Q) and threonyl carbamoyladenosine (t6A). These are two modifications of the tRNA anticodon loop that alter codon definitions and fine tune ribosome function during translation. Q biosynthesis is an established drug target for anti-shigellosis antibiotics. Using a combination of comparative genomic, biochemical, and structural methods, we identified enzymes involved in the biosynthesis of Q and t6A. Using SSRL x-rays, we determined the structure of GCYH-IB, our newly discovered and first enzyme in the Q biosynthesis pathway in prokaryotes, in complex with a substrate analog and we collected native and heavy-atom derivative data for YkvM, the enzyme that catalyzes the third step in Q biosynthesis. We also determined the crystal structure of YrdC, a universally conserved and essential enzyme involved in t6A biosynthesis, in complex with its ATP substrate. In this renewal, we plan to start crystallographic studies of GCYH-IBinhibitor complexes and of QueA, which catalyzes a late step in Q biosynthesis, in complex with its tRNA substrate. We currently have crystals of GCYH-IB-ligand complexes and crystallization of the latter complex is underway. We plan to continue to use the SSRL beamline automation technology and remote access for these studies.

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们已经使用SSRL束时间对这一建议进行晶体学研究的酶负责的生物合成的两个tRNA修饰：queosine（Q）和苏氨酰氨基甲酰基腺苷（t6 A）。这是tRNA反密码子环的两种修饰，它们在翻译过程中改变密码子定义并微调核糖体功能。Q生物合成是抗志贺氏菌抗生素的既定药物靶标。使用比较基因组学，生物化学和结构方法的组合，我们确定了参与Q和t6 A的生物合成的酶。使用SSRL X射线，我们确定了GCYH-IB的结构，GCYH-IB是我们新发现的第一种酶，在原核生物中的Q生物合成途径中，与底物类似物复合，我们收集了YkvM的天然和重原子衍生物数据，YkvM是催化Q生物合成第三步的酶。我们还确定了YrdC的晶体结构，YrdC是一种普遍保守的和必需的酶，参与t6 A的生物合成，与其ATP底物复合。在这次更新中，我们计划开始GCYH-IB抑制剂复合物和QueA的晶体学研究，QueA催化Q生物合成的后期步骤，与其tRNA底物复合。我们目前有GCYH-IB-配体复合物的晶体，后者复合物的结晶正在进行中。我们计划继续使用SSRL光束线自动化技术和远程访问这些研究。