Exploiting New Fibril Structures to Understand the Biophysical Basis for Oligomerization and Toxicity of Alpha-Synuclein

利用新的原纤维结构来了解 α-突触核蛋白寡聚化和毒性的生物物理基础

基本信息

批准号：
10684133
负责人：
Jonathan N Sachs
金额：
$ 37.99万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-30 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10684133
关键词：
Affinity Alzheimer&apos s Disease Amino Acid Motifs Amino Acid Sequence Amino Acids Back Biological Assay Biology Biophysics Catalogs Cell Line Cell model Cells Chemicals Collaborations Color Communities Computer Models Coupled Cytoprotection Data Dementia Disease Event Experimental Designs Fluorescence Fluorescence Resonance Energy Transfer Goals Investigation Kinetics Label Medicine Methodology Microscopy Modeling Molecular Molecular Structure Molecular Weight Monitor Morphologic artifacts Mutation Nature Neurons Neurosciences Outcome Paper Parkinson Disease Pathology Pathway interactions Positioning Attribute Proteins Protocols documentation Publications Recording of previous events Research Research Personnel Resolution Science Series Signal Transduction Structure System Techniques Technology Testing Therapeutic Time Total Internal Reflection Fluorescent Toxic effect Variant Work advanced simulation algorithm development alpha synuclein beta pleated sheet biophysical analysis biophysical tools cell type cytotoxic cytotoxicity design drug discovery high throughput screening inhibitor innovation insight kinetic model molecular modeling molecular scale monomer mutant neuron loss new therapeutic target novel novel strategies prevent protein folding protein misfolding rational design screening small molecule small molecule inhibitor time use tool

项目摘要

Abstract Research into the molecular basis of Parkinson’s Disease has recently undergone a dramatic shift to focus on toxic, early stage oligomers of α-Synuclein (aSyn). Understanding this promising new therapeutic target, a departure from research on insoluble fibrils, now requires biophysical insight about the misfolding of aSyn monomers and subsequent assembly of these toxic oligomers. These oligomer species are far less understood than fibrils, and more difficult to study, presenting a pressing challenge to biophysicists. The specific overall goal of the proposed work is to identify a subset of amino acid interactions within and between aSyn monomers that are most important in the assembly and toxicity of oligomers. Several new high- resolution structures of aSyn fibrils will be used as an exciting starting point to launch detailed investigations into the structural motifs that are present in the early stages of assembly. Based on strong preliminary results, we hypothesize that, despite their relative structural disorder, there exist robust, targetable structural motifs in early stage oligomers that persist through fibrilization. Additionally, a subset of those motifs is essential in determining toxicity: some promote toxic assemblies while others promote cytoprotective assemblies. High-resolution structures of early-stage oligomers will likely never be solved. Absent structures, our data will do the next best thing: it will point to specific motifs and residues that stabilize early-stage oligomers and that should be the focus of directed targeting campaigns. We have established a highly resolved technology (both temporally and spatially), time-resolved FRET, that allows us to study with great sensitivity the early-stages of aSyn aggregation in the cell. We will support these cellular observations with rigorous biophysical studies including 19F NMR, two-color TIRF microscopy and computational modeling. We will also utilize our established small molecule discovery technology in an innovative way to establish whether there are clear structural differences in oligomeric assemblies of the familial variants of aSyn, and whether these assemblies vary in differing neuronal cell lines. In sum, the proposal will provide the field with a significantly deeper understanding of the biophysical basis of aSyn oligomerization and will draw new correlations between key amino-acid residues, folding and toxicity.

抽象的帕金森氏病分子基础的研究最近发生了巨大的转变关于α-突触核蛋白的早期寡聚物（ASYN）。偏离研究原纤维的研究，现在需要生物物理学的见解，以折叠式ASYN 这些低聚物物种的单体和随后的组装。比原纤维和更多的研究，对生物物理学提出了挑战。支撑工作的具体总体目标是确定和内部的氨基酸相互作用的子集在几个新的高级高素质中最重要的Asyn单体之间。 Asyn Fibrils遗嘱的分辨率结构将用于ASED，令人兴奋基于强烈预序的结构基序是在组装的早期阶段。假设尽管存在相对结构性障碍，但早期存在鲁棒，可靶向的结构基序通过纤维化持续存在的低聚物。 tome促进有毒组件，而其他人则促进了细胞保护组件早期低聚物的结构可能永远不会解决。事物：它将指出稳定低聚物的特定图案和残留物定向目标运动。我们已经建立了一项高度解决的技术（在时间和空间上），时间分辨率的货物，该技术是，允许我们以极大的敏感性研究ASYN在细胞中的早期阶段。严格生物物理研究的细胞观察包括19f NMR，两色TIRF显微镜和计算建模。确定此处是否是家族变体的寡聚组件的明显差异 Asyn以及这些组件是否在不同的神经元细胞系中有所不同。总而言之，该提案为该领域提供了对生物物理基础的更深入的了解 Asyn寡聚化，并将在关键氨基酸残基，折叠和毒性之间提出新的相关性。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Advancements in a FRET Biosensor for Live-Cell Fluorescence-Lifetime High-Throughput Screening of Alpha-Synuclein.

DOI：
10.1177/17590914231184086
发表时间：
2023-01
期刊：
ASN NEURO
影响因子：
4.7
作者：
Braun, Anthony R.;Kochen, Noah Nathan;Yuen, Samantha L.;Liao, Elly E.;Cornea, Razvan L.;Thomas, David D.;Sachs, Jonathan N.
通讯作者：
Sachs, Jonathan N.