The molecular determinants of surface-templated self-association of intrinsically disordered proteins

项目摘要 我们研究计划的长期目标是利用新的技术和范例来理解 生理和疾病相关现象的分子决定因素与内在联系 无序蛋白质(IDPs)，即在溶液中不能折叠成稳定结构的蛋白质。我们的关注点是自我 出现在高亲和力表面(如微管或细胞器膜)上的IDPs的结合。表面 可通过增加局部IDP浓度和模板IDP构象更多地促进自缔合 容易自我联想的。然而，国内流离失所者及其各自的高亲和力表面都受到 大量的细胞修饰，极大地扩展了必要的实验参数空间 准确地描述了这一现象。了解自我关联在这个空间中是如何被控制的 对了解它们在生理和疾病中的作用至关重要。因此，我们的实验室将采用新的工具 超越传统的分子生物学，在两个模型系统中重现这种现象 发生：微管上的tau凝聚和突触小泡膜上的α-突触核蛋白多聚体。不 只有PI在这些国内流离失所者的生物物理特征方面拥有广泛的专业知识，但 Tau和α-突触核蛋白为神经生物学和神经退行性疾病提供了丰富的实验历史 可以应用于这一现象的见解。结合起来，这些专业知识和背景可以 纳入了这些境内流离失所者表面模板化的自我联合现象。此外，我们还将 建立易于输出的方案/方法，以研究其他经历表面模板自我模板化的国内流离失所者 协会也是如此。总体而言，我们的意图是通过准确了解与选定的境内流离失所者有关的现象 是创建可推广的机制，以供其他国内流离失所者行为，最终提供一个严格的框架 对这些蛋白质具有解释力和预测力。通过开展拟议的研究，我们希望 将我们的纯生物物理实验室转变为一个完全多学科的项目，将蛋白质的行为 到细胞现象。

Project Summary The long-term goal of our research program is to utilize new techniques and paradigms to understand the molecular determinants of physiological and disease-relevant phenomena associated with intrinsically disordered proteins (IDPs), or proteins that do not fold into stable structures in solution. Our focus is the self- association of IDPs that occur on high-affinity surfaces (such as microtubules or organelle membranes). Surfaces can promote self-association by increasing the local IDP concentration and templating IDP conformations more susceptible to self-association. However, both IDPs and their respective high-affinity surfaces are subject to numerous cellular modifications, dramatically expanding the experimental parameter space necessary to precisely characterize this phenomenon. Understanding how self-association is controlled in this space could be central to understanding their function in physiology and disease. Thus, our laboratory will adapt novel tools beyond traditional molecular biology to recreate conditions in two model systems where this phenomenon occurs: Tau condensation on microtubules and α-synuclein multimerization on synaptic vesicle membranes. Not only does the PI have extensive expertise with biophysically characterizing these IDPs, but the importance of Tau and α-synuclein to neurobiology and neurodegenerative disease provide a rich history of experimental insights that can be applied towards this phenomenon. Combined, this expertise and background can be incorporated into the phenomenon of surface-templated self-association of these IDPs. Furthermore, we will establish protocols/methods that can be easily exported to study other IDPs that undergo surface-templated self- association, as well. Overall, our intention by precisely understanding phenomena associated with select IDPs is to create generalizable mechanisms by which other IDPs behave, eventually providing a rigorous framework that has explanatory and predictive power for these proteins. By undertaking the proposed research, we hope to transition our purely biophysics laboratory to an entirely multidisciplinary program that connects protein behavior to cellular phenomenon.