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技术和范式来了解 生理和疾病相关现象的分子决定因素， 无序蛋白（IDP）或在溶液中不能折叠成稳定结构的蛋白质。我们关注的是自我- 发生在高亲和力表面（如微管或细胞器膜）上的IDP的缔合。表面 可以通过增加局部IDP浓度和模板IDP构象来促进自缔合 容易自我联想。然而，IDP和它们各自的高亲和力表面都受到 大量的细胞修饰，极大地扩展了实验参数空间， 准确地描述了这种现象。了解自我联想在这个空间中是如何控制的， 对了解它们在生理学和疾病中的功能至关重要。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凝聚和突触囊泡膜上的α-突触核蛋白多聚化。不 只有PI具有广泛的专业知识，可以对这些IDP进行生物识别，但重要的是 Tau和α-synuclein为神经生物学和神经退行性疾病提供了丰富的实验研究历史 可以应用于这种现象的见解。结合起来，这种专业知识和背景可以 被纳入这些国内流离失所者的表面模板化自我关联现象。此外，我们将 建立协议/方法，可以很容易地出口到研究其他IDP经历表面模板化的自我， 协会，也是。总的来说，我们的意图是准确理解与特定国内流离失所者有关的现象， 是为其他国内流离失所者的行为建立一个可推广的机制，最终提供一个严格的框架 对这些蛋白质有解释和预测能力的。通过开展拟议的研究，我们希望 将我们纯粹的生物物理实验室转变为一个完全多学科的项目， 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.