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.

项目概要/摘要 现在人们对蛋白质折叠有了更好的了解，我们可以将其与其他相互作用结合起来研究 蛋白质与生物分子形成的。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，例如RNA。它们还用聚乙二醇 (PEG) 等附件进行修饰 可以增强药物输送稳定性的分子。最后，较大蛋白质的结构域可以与一种蛋白质相互作用 另一种是改变折叠过程或导致不需要的聚集，这可能会导致蛋白质 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，并 定量地描述这些相互作用。在这一长期目标中，我们的具体目标有三个： 1) PEG广泛应用于制药行业，以改善蛋白质药物的递送。我们学习 PEG 如何与蛋白质表面相互作用，因此我们可以找出 PEG 帮助稳定的机制 用于递送的蛋白质药物。我们将研究几种蛋白质系统，包括慢性病的治疗剂 肾脏疾病。 2）剪接体在细胞核中组装，剪接并重新组装信使RNA，即 获取从细胞核到核糖体制造新蛋白质的信息是必要的，蛋白质在核糖体中 合成的。我们将通过制作许多突变体并进行比较来研究关键的蛋白质-RNA 相互作用之一 他们使用我们刚刚开发的新模型，我们认为该模型可以预测蛋白质和 RNA 的结合强度。 这对于合理设计干扰或修复蛋白质-RNA 相互作用的药物非常重要。 3）大蛋白质包含许多结构域，当它们折叠时可能会出错。我们研究这些相互作用 在压力和温度的扩展相图中，更好地了解它们的物理起源。我们 发现折叠中间体（即未完全折叠的结构）可能会出现并 消失在该相图中。通过了解为什么会发生这种情况，我们可以更好地抑制此类中间体， 这可能会形成有害的聚集体。 为了实现我们的目标，我们正在开发新的荧光检测方法，以快速、灵敏地检测 互动。我们正在扩展蛋白质加压技术的能力，因此我们可以研究 与食品压力灭菌相关条件下的蛋白质。我们正在制作新的 PEG 标记的 蛋白质来研究 PEG 长度和附着位点的真正重要性。