Oxytocin Modulation of Social Behavior

项目概要/摘要： 这个提议的目的是剖析围绕成核和早期事件的机制细节 tau蛋白聚集Tau蛋白是一种关键的错误折叠和聚集蛋白， (AD)额颞叶痴呆（FTD）在过去的几年里， 患有神经退行性疾病，包括AD和FTD，预计将继续上升， 目前还没有治愈方法。虽然许多研究将tau蛋白的聚集归因于该疾病的原因， tau聚集的成核和自分裂过程还没有被很好地理解。一个主要挑战是 已经确定了tau错误折叠和聚集的构象基础，由于大尺寸的 蛋白为了解决这一挑战，31个残基的最小序列已被定义为构象 核负责tau聚集的自我传播，从而为研究 成核的早期阶段。根据这一观察结果，我们假设在这个最小序列中的残基 与蛋白质的其他结构域相互作用以诱导单分子水平的聚集以及相互作用 与其他错误折叠的tau分子一起引发聚集。为了验证这一点，我们建议将这些相互作用映射到一个 位点特异性方式使用最小干扰的荧光非天然氨基酸来阐明 非纤维状低聚物形成和聚集过程中的局部构象动力学。 创新：为了剖析位点特异性相互作用，动力学和构象动力学周围的 成核和τ的早期聚集事件，需要新的工具。非天然氨基酸对 水合作用对于探测聚集是唯一有用的，因为当 一旦可溶性蛋白质在聚集过程中变得不溶。目前的技术，如 荧光蛋白和染料由于它们的尺寸过大而不适合于研究分子内事件。不自然 氨基酸发色团在一个框架中提供所需的光谱性质， tau的结构和功能，使其成为研究分子内和分子间构象的理想选择。 动力学、动力学和tau聚集过程中的相互作用。这将使我们的直接知识， 有助于阐明聚集途径。

Project Summary/Abstract: The Goal of this proposal is to dissect the mechanistic details surrounding the nucleation and early events of tau aggregation. Tau is a key misfolding and aggregating protein associated with both Alzheimer’s disease (AD) and frontotemporal dementia (FTD). There has been a significant increase in the number of people suffering from neurodegenerative diseases, including AD and FTD and is expected to continue to rise as there are currently no cures. While many studies have attributed the aggregation of tau as a cause of the disease, the process of nucleation and self-prorogation of tau aggregation is not well understood. One major challenge has been to determine the conformational basis of tau misfolding and aggregation due to the large size of the protein. To address this challenge, minimal sequence of 31 residues has been defined as the conformational nucleus responsible for the self-propagation of tau aggregation, thus providing a foundation for investigating the early stage of nucleation. With this observation we hypothesize that residues within this minimal sequence interact with other domains of the protein to induce aggregation at the single molecule level as well as interact with other misfolded tau molecules to seed aggregation. To test this, we propose to map these interactions in a site specific manner using minimally perturbing fluorescent unnatural amino acids to shed light on the early non-fibrillar oligomer formation and the local conformational dynamics over the course of aggregation. Innovation: In order to dissect site-specific interactions, kinetics and conformational dynamics surrounding the nucleation and early aggregation events of tau, new tools are needed. Unnatural amino acids sensitive to hydration are uniquely useful to probe aggregation because of the change in local hydration exhibited when once soluble protein becomes insoluble during aggregation. Unfortunately, current technologies such as fluorescent proteins and dyes are not ideal to study intramolecular events due to their prohibitive size. Unnatural amino acids chromophores offer the desired spectroscopic qualities in a framework that is non-perturbing to both structure and function of tau, making it ideal to study the intra- and intermolecular conformational dynamics, kinetics and interactions during tau aggregation. Together this will inform our direct knowledge to help elucidate the aggregation pathway.