Protein domains interacting with crowders, RNA and other protein domains

与 Crowder、RNA 和其他蛋白质结构域相互作用的蛋白质结构域

• 批准号: 9372464
• 负责人: MARTIN GRUEBELE
• 金额: $ 29.24万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9372464
• 关键词: Affect; Affinity; Amyloid; Back; Binding; Binding Proteins; Biological Assay; Biological Models; Biophysics; Categories; Cell Nucleus; Cells; Charge; Chronic Kidney Failure; Communities; Complement; Complex; Computers; Coupled; Crowding; Data; Disease; Drug Delivery Systems; Drug Design; Drug Industry; Drug Stability; Drug Targeting; Electrostatics; Equilibrium; Fluorescence; Fluorescence Resonance Energy Transfer; Future; Goals; Health; Human Resources; Hydrogen Bonding; Hydrophobic Interactions; Illinois; Industry; Kinetics; Knowledge; Label; Lead; Learning; Length; Measurement; Measures; Medicine; Membrane Proteins; Messenger RNA; Modeling; Names; Organism; Patients; Pharmaceutical Preparations; Phase; Phosphoglycerate Kinase; Physiological; Polyethylene Glycols; Polymers; Population; Price; Problem Solving; Process; Protein Binding Domain; Protein Dynamics; Proteins; Proteolysis; Publishing; RNA; RNA Binding; RNA Splicing; RNA-Binding Proteins; RNA-Protein Interaction; Research; Ribosomal Proteins; Scanning; Science; Scientist; Side; Site; Site-Directed Mutagenesis; Spliceosomes; Structure; System; Techniques; Temperature; Tertiary Protein Structure; Testing; Therapeutic Agents; Thermodynamics; Universities; base; design; experimental study; fascinate; food sterilization; improved; in vitro Assay; lambda repressor; migration; mutant; new therapeutic target; predictive modeling; pressure; protein folding; protein protein interaction; repaired; simulation; small molecule; theories; therapeutic protein

Project Summary/Abstract Now that protein folding is becoming better understood, we can study it in combination with other interactions that proteins make with biomolecules. Proteins in cells and organisms continuously interact with other biomolecules, such as RNA. They are also modified with attachments, such as polyethylene glycol (PEG) molecules that can enhance stability for drug delivery. Finally, domains of larger proteins can interact with one another, modifying the folding process or leading to undesirable aggregation, which can lead to protein diseases. Our long-term objective is to study interactions of proteins with PEG, RNA, and other protein domains, and to characterize these interactions quantitatively. Within that long-term objective, our specific aims are threefold: 1) PEG is used extensively in the pharmaceutical industry to improve the delivery of protein drugs. We study how PEG interacts with protein surfaces, so we can figure out the mechanism by which PEG helps stabilize protein drugs for delivery. We will study several protein systems, including a therapeutic agent for chronic kidney disease. 2) The spliceosome assembles in the cell nucleus to splice and re-assemble messenger RNA, which is necessary to take the information to make new proteins from the nucleus to the ribosomes, where proteins are synthesized. We will study one of the key protein-RNA interactions by making many mutants and comparing them with a new model we just developed, that we think can predict how strongly protein and RNA will bind. This will be important for rational design of drugs to interfere with, or repair, protein-RNA interactions. 3) Large proteins contain many domains, and when they fold things can go wrong. We study these interactions in an expanded phase diagram of pressure and temperature, to better understand their physical origins. We discovered that folding intermediates, which are structures that are not quite properly folded, can appear and disappear in this phase diagram. By learning why this happens we can better suppress such intermediates, which could form harmful aggregates. To achieve our goals, we are developing new fluorescence assays to rapidly and sensitively detect interactions. We are expanding the capabilities of our protein pressurization techniques, so we can study protein under conditions relevant to pressure sterilization of food. And we are making new PEG-labeled proteins to study how important PEG length and attachment sites really are.

项目总结/摘要 既然蛋白质折叠已经被更好地理解，我们可以结合其他相互作用来研究它 蛋白质与生物分子的结合。细胞和生物体中的蛋白质不断地与其他蛋白质相互作用， 生物分子，如RNA。它们也被修饰了附件，如聚乙二醇（PEG） 这些分子可以增强药物输送的稳定性。最后，较大蛋白质的结构域可以与 另一个，改变折叠过程或导致不希望的聚集，这可能导致蛋白质 疾病 我们的长期目标是研究蛋白质与PEG、RNA和其他蛋白质结构域的相互作用， 定量地描述这些相互作用。在这一长期目标范围内，我们的具体目标有三个方面： 1)PEG在制药工业中广泛用于改善蛋白质药物的递送。我们研究 PEG如何与蛋白质表面相互作用，因此我们可以找出PEG帮助稳定蛋白质的机制， 蛋白质药物的递送。我们将研究几种蛋白质系统，包括一种治疗慢性 肾病 2)剪接体在细胞核中组装以剪接和重新组装信使RNA， 从细胞核到核糖体，蛋白质在核糖体中的位置， 合成了我们将通过制造许多突变体和比较来研究关键蛋白质-RNA相互作用之一 用我们刚刚开发的一个新模型，我们认为这个模型可以预测蛋白质和RNA结合的强度。 这对于合理设计药物来干扰或修复蛋白质-RNA相互作用至关重要。 3)大型蛋白质包含许多结构域，当它们折叠时，事情可能会出错。我们研究这些相互作用 在压力和温度的扩展相图中，以更好地理解它们的物理起源。我们 发现折叠中间体，即没有完全折叠的结构，可以出现， 在这个相图中消失。通过了解为什么会发生这种情况，我们可以更好地抑制这些中间体， 这可能形成有害的聚集体。 为了实现我们的目标，我们正在开发新的荧光检测方法， 交互.我们正在扩大蛋白质加压技术的能力， 在与食品压力灭菌相关的条件下，我们正在制造新的PEG标记的 研究PEG长度和附着位点的重要性。