STRUCTURES OF SIGNALING COMPLEXES INVOLVED IN BACTERIAL CHEMOTAXIS

参与细菌趋化性的信号复合物的结构

• 批准号: 7955540
• 负责人: BRIAN R CRANE
• 金额: $ 4.89万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955540
• 关键词: Azurin; Chemotaxis; Complex; Computer Retrieval of Information on Scientific Projects Database; Copper; Cytochrome c Peroxidase; Data; Electron Transport; Enzymes; Equus caballus; Funding; Goals; Grant; Heart; Indoles; Institution; Maps; Measures; Metals; Molecular Conformation; Porphyrins; Procollagen-Proline Dioxygenase; Proteins; Reaction; Research; Research Personnel; Resolution; Resources; Signal Transduction; Site; Solutions; Solvents; Source; Structure; System; Time; Tryptophan; United States National Institutes of Health; Yeasts; base; cofactor; cytochrome c; electron density; improved; mutant; programs; protein structure

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. A central goal of our program is to understand how protein structure controls long-range electron transfer (ET) reactions. Three systems were studied this past year - 1) Interprotein ET in complexes of cytochrome c (Cc) with cytochrome c peroxidase (CcP) and 2) Inter- and intra protein ET reactions in metal-modified azurins (Az) 3) Inter-cofactor ET in prolyl 4-hydroxylase (P4H). 1)CcP:Cc - We determined structures of Zn-porphyrin containing CcP with Cc from yeast (yCc) and Cc from horse heart (hCc). YCc and hCc have different modes of interaction with CcP. Electron transfer rates measured directly in these crystals confirm for the first time that the crystal structures represent associations that govern solution reactivity. Moreover ET rate calculations based these new structures indicate that the observed reactivity can only be rationalized if conformational change gates ET. We have also collected diffraction data on complexes that contain site-directed mutants of Cc that drastically alter electron transfer reactivity. 2) Az - We determined the high resolution structure of a Re-modified azurin in which a very stable tryptophan radical forms by photochemically triggered oxidative electron transfer to the Re. Surprisingly, the Trp site of the stable radical is solvent exposed and the Trp indole is found in multiple conformations. We have also determined an atomic resolution structure (0.98 ¿ resolution) of a Cu and Zn-containing azurin. These are the first atomic resolution structures of a blue-copper protein. 3) P4H - We have progressed towards determining the first structure for a collegen-type P4H, an enzyme where inter-cofactor ET rescues oxidatively trapped states. We have now collected diffraction data on three different crystal forms and have improved our native data to 2.4 ¿ resolution. We have collected anomalous scattering data from Pb and W derivatives and have an interpretable electron density map. Structure determination is underway.

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。 我们程序的一个核心目标是了解蛋白质结构如何控制远程电子转移（ET）反应。在过去的一年中，研究了三个系统 - 1）在金属改性偶氮（AZ）3）中的细胞色素C（CC）中与细胞色素C过氧化物酶（CCP）（CCP）（CCP）和2）蛋白质蛋白ET反应（Az）3）中型蛋白ET反应中的分解蛋白ET进行了研究。 1）CCP：CC-我们确定了含有酵母（YCC）CC的Zn -Porphyrin的结构，从马心（HCC）进行CC。 YCC和HCC与CCP具有不同的相互作用模式。直接在这些晶体中测量的电子转移速率首次证实晶体结构代表了控制溶液反应性的关联。此外，基于这些新结构的ET速率计算表明，只有在会议变更门等的情况下，观察到的反应性才能合理化。我们还收集了包含CC位置导向突变体的复合物的衍射数据，这些突变体极大地改变了电子传递反应性。 2）AZ-我们确定了重新修饰的蓝蛋白的高分辨率结构，其中通过光化学触发的氧化电子转移到RE，一种非常稳定的色氨酸自由基形式。令人惊讶的是，稳定自由基的TRP位点被溶解了暴露，并且在多个考虑因素中发现了TRP吲哚。我们还确定了含Cu和Zn的硫蛋白的原子分辨率结构（0.98»分辨率）。这些是蓝opper蛋白的第一个原子分辨率结构。 3）P4H-我们已经进步朝着确定collegen型P4H的第一个结构，这是一种酶，该酶响应氧化性捕获的状态。现在，我们已经在三种不同的晶体形式上收集了衍射数据，并将本机数据提高到2.4`分辨率。我们从PB和W衍生物中收集了异常的散射数据，并具有可解释的电子密度图。结构确定正在进行中。