Structural and mechanistic studies of PAS sensing

PAS传感的结构和机制研究

• 批准号: 10436974
• 负责人: Kevin H Gardner
• 金额: $ 36.11万
• 依托单位: ADVANCED SCIENCE RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10436974
• 关键词: ARNT gene; Address; Attention; Bacteria; Biochemical; Biological; Biological Assay; Biology; Biomedical Research; Biophysics; Biotechnology; Cell membrane; Cell physiology; Cells; Characteristics; Chemicals; Coupled; Coupling; DNA Binding; Data; Development; Disease; Engineering; Environment; Enzymes; Event; Family; Family member; Flavins; Foundations; GTP-Binding Protein Regulators; GTP-Binding Proteins; Gene Expression; Generations; Genomics; Goals; Helix-Turn-Helix Motifs; Human; In Vitro; Left; Length; Ligands; Light; Lighting; Membrane; Modeling; Molecular Conformation; Nature; Nutrient; Organism; Outcome; Outcomes Research; Output; Oxygen; Pathway interactions; Phosphotransferases; Process; Protein Binding Domain; Protein Conformation; Protein-Serine-Threonine Kinases; Proteins; RGS Domain; Reagent; Regulation; Research; Research Personnel; Resolution; Route; Sensory; Signal Transduction; Signaling Protein; Stimulus; Structure; Structure-Activity Relationship; System; Techniques; Tertiary Protein Structure; Testing; Therapeutic; Work; Xenobiotics; Yeasts; adduct; anti-cancer therapeutic; base; chromophore; cofactor; detection of nutrient; environmental change; experimental study; fungus; glucose uptake; innovation; insight; novel; novel therapeutics; optogenetics; protein function; protein-histidine kinase; recruit; response; sensory mechanism; small molecule; success; therapeutic target; tool; transcription factor; voltage

ABSTRACT For cells to properly adapt to changing environmental conditions around them, they must rely on sensory proteins which both detect these changes and initiate proper responses. As one solution to this challenge, nature has evolved classes of ligand-regulated protein/protein interaction domains, harnessing these to control numerous types of protein function. Understanding how these domains undertake the requisite sensing and signaling events has given insight into fundamental aspects of biology, enabled the development of new tools for biomedical research, and inspired novel therapies to disease. Here we focus on examining the signaling mechanisms used by proteins containing PAS (Period-ARNT-Singleminded) domains, found in tens of thousands of proteins where they control the activity of over 20 enzymatic and non-enzymatic effector domains. These processes are regulated by different stimuli in different members of the family, harnessing changes in the occupancy or configuration of bound cofactors to “switch” activity on and off. Some aspects of these triggers are well understood in some PAS domains, such as in the subset known as LOV (Light-Oxygen- Voltage) photosensory domains, where blue light illumination drives the specific photochemical formation of protein/flavin adducts which allosterically controls protein conformation around the chromophore. However, fundamental questions regarding these signaling processes remain unanswered, limiting our understanding of how environmental changes are sensed and how these might be artificially controlled. We propose to answer these limitations by pursuing three aims: 1). Determine activation-associated structural changes of several classes of bacterial LOV and PAS proteins; 2). Examine the generality of LOV signaling within a novel class of fungal RGS-LOV proteins; 3). Examine PAS regulation of serine/threonine kinase activity in the nutrient- sensing human PAS kinase. To achieve these ends, we will take advantage of a broad foundation of preliminary structural and functional data that will be extended with a mix of biophysical and biochemical studies. Outcomes from this research will include information about fundamental regulatory processes employed by these proteins, giving insights into both basic aspects of biomedicine along with potential therapeutic and biotechnology applications.

摘要 细胞要想适当地适应周围不断变化的环境条件，就必须依靠感觉。 既能检测到这些变化又能引发适当反应的蛋白质。作为这一挑战的一种解决方案， 自然界进化出了一类受配体调节的蛋白质/蛋白质相互作用结构域，利用这些结构域来控制 蛋白质的功能种类繁多。了解这些域如何承担必要的感知和 信号事件使人们深入了解生物学的基本方面，使新工具的开发成为可能 用于生物医学研究，并启发了疾病的新疗法。在这里，我们重点检查信令 含有PAS(Period-Arnt-Singlminded)结构域的蛋白质所使用的机制，在数十个 数以千计的蛋白质，它们控制着20多种酶和非酶效应器的活性 域名。这些过程在不同的家庭成员中受到不同刺激的调节，驾驭 改变结合辅因子的占有率或配置，以“开启和关闭”活动。在某些方面 这些触发器在一些PAS域中被很好地理解，例如在被称为LOV(光-氧- 电压)光敏感区，其中蓝光照明驱动特定的光化学形成 蛋白质/黄素加合物以变构方式控制发色团周围的蛋白质构象。然而， 关于这些信令过程的基本问题仍然没有得到回答，限制了我们对 环境变化是如何被感知的，以及这些变化可能如何被人为控制。我们建议回答 这些局限通过追求三个目标来实现：1)。确定几个与激活相关的结构变化 细菌LOV和PAS蛋白的分类；研究LOV信令在一类新的 真菌RGS-LOV蛋白；检测PAS对营养物质中丝氨酸/苏氨酸激酶活性的调节 感应到人的PAS激酶。为了达到这些目标，我们将利用 初步的结构和功能数据，将与生物物理和生化的混合进行扩展 学习。这项研究的结果将包括有关基本监管过程的信息 被这些蛋白质利用，使人们对生物医学的两个基本方面以及潜在的 治疗和生物技术应用。