HIGH-RESOLUTION CRYSTALLOGRAPHIC STUDIES OF TWO RESPIRATORY PROTEINS FROM T THE

两种呼吸蛋白的高分辨率晶体学研究

• 批准号: 7722120
• 负责人: JAMES A FEE
• 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/7722120
• 关键词: Binding; Biochemistry; Computer Retrieval of Information on Scientific Projects Database; Condition; Crystallization; Enzymes; Funding; Grant; Heme; Imidazole; In Situ; Institution; Ligation; Mutate; Mutation; Oxidases; Proteins; Protons; Research; Research Personnel; Resolution; Resources; Roentgen Rays; Screening procedure; Source; Structure; System; Thermus; Time; United States National Institutes of Health; Work; cryogenics; cytochrome c oxidase; fluoromethyl 2,2-difluoro-1-(trifluoromethyl)vinyl ether; insight; interest; mutant; novel; oxidation; respiratory protein

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. In order to garner powerful new insights on heme-Cu oxidase biochemistry, continued X-ray beam time is requested to perform crystallographic studies of the novel cytochrome c oxidase, ba3, from T. thermophilus. Our plans call for obtaining: (a) increased resolution of a new mutant form of the as-isolated, fully oxidized enzyme; (b) independent structures of numerous mutant forms notably: (i) of mutated proteins that crystallize in different space groups with the view of increasing reliability of crystallization in different oxidation and ligation states, and, we hope, through rational screening, will provide ~2.0 A resolution, (ii) to reveal both structure and function of the O2 delivery channel, (iii) to examine the structural effects of mutations in the so-called K-path of proton transport; (c) independent structures of the fully reduced enzyme followed by in situ oxygenation with gaseous O2 using a cryogenic flow system under conditions which permit formation of the cryo-stable A-compound, Fea3-O2. This constitutes the main objectives of our effort. However, we retain an interest in the Thermus Rieske protein. Previous studies yielded a 1.3 ¿ structure of the fully-oxidized Thermus Rieske protein at pH 8.5 in which one of the Fe-bound imidazole rings was deduced to be partially de-protonated. Our plans call for extending this work to include the following: (a) an atomic resolution (

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 为了获得有关Heme-Cu氧化酶生物化学的强大新见解，请求继续X射线束时间，以对T. thermophilus进行新型细胞色素C氧化酶BA3进行晶体学研究。我们的计划呼吁获得：（a）增加了新型突变体形式的分离，完全氧化的酶； (b) independent structures of numerous mutant forms notably: (i) of mutated proteins that crystallize in different space groups with the view of increasing Reliability of crystallization in different oxidation and ligation states, and, we hope, through rational screening, will provide ~2.0 A resolution, (ii) to reveal both structure and function of the O2 delivery channel, (iii) to examine the structural effects of mutations in the so-called K-path of proton transport; （c）在允许形成冷冻稳定的A-Compound FEA3-O2的条件下，使用气态O2使用气态O2的原位氧合的独立结构，然后使用气态O2进行原位氧合。这构成了我们努力的主要目标。但是，我们保留对Thermus Rieske蛋白的兴趣。先前的研究产生了pH 8.5的全氧化热雷脉蛋白的结构1.3€，其中一种被推断为部分去蛋白酶对照的Fe-bond-BOND结合的环。我们的计划要求将此工作扩展到以下内容：（a）原子分辨率（