Protein domains interacting with crowders, RNA and other protein domains

项目摘要/摘要 现在人们对蛋白质折叠有了更好的理解，我们可以结合其他相互作用来研究它 蛋白质用生物分子制造的物质。细胞和生物体中的蛋白质不断地与其他 生物分子，如核糖核酸。它们还可以用附件进行修饰，如聚乙二醇酯(PEG)。 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个蛋白质相互作用。 另一种是改变折叠过程或导致不必要的聚集，这可能导致蛋白质 疾病。 我们的长期目标是研究蛋白质与聚乙二醇、核糖核酸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方面： 1)在制药工业中广泛使用聚乙二醇酯，以改善蛋白质药物的输送。我们学习 聚乙二醇如何与蛋白质表面相互作用，这样我们就可以弄清楚聚乙二醇帮助稳定的机制 用于运送的蛋白质药物。我们将研究几种蛋白质系统，包括一种治疗慢性阻塞性肺疾病的药物 肾脏疾病。 2)剪接体在细胞核内组装，剪接和重组信使RNA，这是 从细胞核到核糖体产生新的蛋白质所需的信息，在那里蛋白质 合成的。我们将通过制造许多突变体和比较来研究蛋白质-RNA相互作用的关键之一 我们刚刚开发了一个新的模型，我们认为该模型可以预测蛋白质和RNA结合的强度。 这对于合理设计干扰或修复蛋白质-RNA相互作用的药物将是重要的。 3)大的蛋白质包含许多结构域，当它们折叠时，事情可能会出错。我们研究了这些相互作用 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，这是不完全折叠的结构，可以出现和 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， 它们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分析方法，以快速和灵敏地检测 互动。我们正在扩展我们的蛋白质加压技术的能力，这样我们就可以研究 与食品加压灭菌相关的条件下的蛋白质。我们正在制作一种新的聚乙二醇式标签 蛋白质来研究聚乙二醇链长度和连接位置到底有多重要。

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.