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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 所有生物体都有能力对生物刺激做出反应，例如光电压、氧配体和环境因素。当施加时，每种刺激在几个步骤中转化为特定的细胞反应，从而促进信号转导途径。为了理解信号转导的工作机制，重要的是要识别参与信号传导的分子单位，例如刺激受体。 在刺激中，光在广泛的生物学重要细胞反应中起着重要作用，包括光合作用和基因表达（1）。到目前为止，已经确定了六种光感受器（2）。在这些光感受器中，BLUF结构域是最近发现的，因此已经在有限程度上进行了研究。特别地，Slr 1694 BLUF结构域在溶液中作为同源二聚体存在，但在黑暗中寡聚形成十聚体。这种推定的信号传导状态是光学惰性的，其形成动力学不容易使用时间分辨光谱学例如瞬态吸收（TA）光谱学来探测。相反，时间分辨X射线散射技术对结构变化敏感，而不管感兴趣状态的光学活性如何。例如，时间分辨小角X射线散射（TR-SAXS）对溶液中蛋白质的大小和形状等全局结构变化敏感，而时间分辨广角X射线散射（TR-WAXS）提供了有关蛋白质三级和四级结构变化的丰富信息。具有全局灵敏度的结构变化。因此，时间分辨X射线散射技术适合于探测BLUF结构域蛋白中信号状态形成的结构动力学。在这里，我们提出的时间分辨实验的Slr 1694 BLUF结构域蛋白使用TR-SAXS和TR-WAXS技术来研究光化学反应导致的信号状态的形成的机制。