SIGNALING STATE OLIGOMERIZATION OF THE SLR1694 BLUF DOMAIN DURING PHOTOCYCLE

光循环期间 SLR1694 BLUF 结构域的信号状态寡聚化

• 批准号: 8363701
• 负责人: HYOTCHERL IHEE
• 金额: $ 9.73万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363701
• 关键词: Biological; Environmental Risk Factor; Funding; Gene Expression; Grant; Homo; Life; Ligands; Light; Molecular; National Center for Research Resources; Optics; Organism; Oxygen; Photoreceptors; Photosynthesis; Play; Principal Investigator; Proteins; Reaction; Research; Research Infrastructure; Resources; Roentgen Rays; Role; Shapes; Signal Transduction; Signal Transduction Pathway; Solutions; Source; Spectrum Analysis; State Interests; Stimulus; Techniques; Tertiary Protein Structure; Time; United States National Institutes of Health; Work; absorption; cost; dimer; receptor; research study; response; structural biology; time use; voltage

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. All living organisms have an ability to respond to biological stimuli such as light voltage oxygen ligands and environmental factors. When applied each stimulus is converted into a specific cellular response in several steps contributing to signal-transduction pathway. To understand the working mechanism of signal transduction it is important to identify the molecular units involved in signaling for example stimuli receptors. Among the stimuli light plays an important role in a wide range of biologically important cellular responses including photosynthesis and gene expression (1). Thus far six kinds of photoreceptors have been identified (2). Among these photoreceptors the BLUF domain was discovered recently thus having been studied to a limited extent. In particular the Slr1694 BLUF domain exists as a homo dimer in solution but oligomerizes in the dark to form a decamer. This putative signaling state is optically inactive that its formation dynamics is not readily probed using time-resolved optical spectroscopy for example transient absorption (TA) spectroscopy. In contrast time-resolved X-ray scattering techniques are sensitive to the structural changes regardless of the optical activity of the state of interest. For example time-resolved small-angle X-ray scattering (TR-SAXS) is sensitive to global structural changes such as size and shape of a protein in solution while time-resolved wide-angle X-ray scattering (TR-WAXS) provides rich information about tertiary and quaternary structural changes of a protein with global sensitivity. Therefore time-resolved X-ray scattering techniques are suitable for probing the structural dynamics of the signaling state formation in the BLUF domain proteins. Here we propose the time-resolved experiments on the Slr1694 BLUF domain proteins using TR-SAXS and TR-WAXS techniques to investigate the mechanism of photochemical reaction leading to the formation of signaling state.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 所有生物体都有能力对生物学刺激做出反应，例如光电压氧配体和环境因素。当应用时，每种刺激被转换为特定的细胞反应，以几个步骤有助于信号变形途径。要了解信号转导的工作机理，重要的是要确定涉及信号传导的分子单位，例如刺激受体。 在刺激中，光在包括光合作用和基因表达在内的广泛的生物学重要细胞反应中起着重要作用（1）。到目前为止，已经鉴定出六种感光体（2）。在这些光感受器中，最近发现了BLUF结构域，因此在有限的程度上进行了研究。特别是SLR1694 BLUF域作为溶液中的同型二聚体存在，但在黑暗中寡聚以形成decamer。这种推定的信号传导状态在光学上是无效的，即使用时间分辨的光谱法（例如瞬时吸收（TA）光谱）不容易探测其形成动力学。相比之下，不论感兴趣状态的光学活性如何，相反的时间分辨X射线散射技术对结构变化敏感。例如，时间分辨的小角度X射线散射（TR-SAXS）对溶液中蛋白质的大小和蛋白质的大小和形状敏感，而时间分解的广角X射线散射（TR-WAXS）则提供了有关具有全球敏感性的蛋白质的第三纪和四级结构变化的丰富信息。因此，时间分辨X射线散射技术适合探测BLUF结构蛋白中信号态形成的结构动力学。在这里，我们建议使用TR-SAXS和TR-WAXS技术在SLR1694 BLUF结构蛋白上进行时间分辨实验，以研究光化学反应的机理，从而导致信号传导的形成。