Understanding and targeting the Methionine-Aromatic motif in oxidized alpha-Synuclein

了解和靶向氧化 α-突触核蛋白中的甲硫氨酸-芳香族基序

基本信息

批准号：
9791033
负责人：
Jonathan N Sachs
金额：
$ 17.64万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-27 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9791033
关键词：
Amino Acids Aromatic Amino Acids Aromatic Compounds Biological Assay Biology Biophysics Biosensor C-terminal Calmodulin Cell Death Cells Chemicals Chemistry Clinic Collaborations Communication Computer Simulation Computers Coupled Data Detection Disease Dopamine Engineering Enzymes Fluorescence Fluorescence Resonance Energy Transfer Future Genetic Transcription Goals Interruption Knowledge Lead Letters Libraries Ligand Binding Ligands Literature Measurement Mediating Methionine Methods Modification Molecular Monitor Nature Nerve Degeneration Neurons Neurosciences Oxidative Stress Oxides Paper Parkinson Disease Pharmaceutical Chemistry Pharmaceutical Preparations Play Process Protein Conformation Proteins Protocols documentation Publishing Reagent Recombinant Proteins Recording of previous events Research Role Sampling Series Side Stretching Structure Sulfoxide Tertiary Protein Structure Testing Thermodynamics Time Toxic effect Tumor Necrosis Factor Receptor Tyrosine Work alpha synuclein amyloid formation base driving force drug discovery effective therapy efficacy testing experimental study fighting high throughput screening inhibitor/antagonist insight methionine sulfoxide molecular dynamics monomer neurotoxic new therapeutic target novel novel therapeutics oxidation prevent protein folding screening simulation small molecule small molecule inhibitor targeted treatment tool

项目摘要

ABSTRACT A distinct feature of Parkinson’s Disease (PD) is the aggregation of neurotoxic α-Synuclein (αSyn) into amyloid assemblies. Despite over two decades of intense efforts to identify inhibitors that directly interrupt the αSyn aggregation process, no successful compounds have reached the clinic. Research on the molecular basis of PD has recently undergone a dramatic shift to focus on toxic oligomeric αSyn and its relation to neurodegeneration. Understanding this promising new therapeutic target, a departure from research on insoluble fibrils, now requires biophysical insight about the misfolding of αSyn monomers into the toxic oligomeric forms of the protein. Several recent studies have pointed to oxidative stress conditions as leading to the formation of highly toxic αSyn oligomers. Thus, our first goal is to understand the molecular basis for misfolding of oxidized αSyn. We will build on several high-impact discoveries made in the past two years that highlighted the importance of C-terminal tyrosine residues in the misfolding process. We will test a straightforward hypothesis that is based on our own recent discovery (published in Nature Chemical Biology in 2016), namely that oxidation of methionine leads to the formation of a strong non-covalent interaction with aromatic residues, including tyrosine. Our approach will provide quantitative details of the chemistry and biophysics of αSyn, including state- of-the-art NMR measurements and computational modeling. Second, we will discover a novel set of small molecules that, for the first time, target the formation and stability of toxic, oxidized αSyn oligomers. We will take advantage of a powerful new platform for high-throughput screening that is based on exquisitely sensitive fluorescence lifetime measurements, and test the functional efficacy of Hit compounds in neurons. These small molecules will ultimately provide a platform for our groups, in future R01-scale proposals, to launch a full-scale medicinal chemistry drug discovery campaign; but in this proposal, the Hit compounds will serve as probes to further determine (by NMR) which specific side-chain interactions with oxidized methionine drive misfolding of αSyn. !

抽象的帕金森氏病（PD）的一个独特特征是神经毒性α-突触核蛋白（αSyn）的聚集成淀粉样蛋白集会。尽管超过二十年的激烈努力鉴定直接中断αSyn的抑制剂聚集过程，没有成功的化合物到达诊所。 PD分子基础的研究最近已经发生了巨大的转变，以关注有毒的寡聚αSyn及其与之的关系神经变性。了解这个有希望的新治疗靶标，与不溶性的研究背道而驰原纤维，现在需要关于αsyn单体错误折叠到有毒寡聚形式的生物物理洞察力最近的一些研究指出，氧化应激条件是导致形成的剧毒αSyn低聚物。这，我们的第一个目标是了解错误折叠的分子基础氧化的αSyn。我们将基于过去两年中的几个高影响力发现，突出了 C末端酪氨酸的重要性在错误折叠过程中保留。我们将检验一个直接的假设这是基于我们自己最近的发现（2016年在自然化学生物学上发表），即氧化蛋氨酸导致形成与芳族保留的强烈非共价相互作用，包括酪氨酸。我们的方法将提供αSyn的化学和生物物理学的定量细节，包括状态 ART NMR测量和计算建模。其次，我们将发现一组新颖的小该分子首次针对有毒，氧化αSyn低聚物的形成和稳定性。我们将利用一个强大的新平台进行高通量筛选，这是基于敏感的荧光寿命测量值，并测试神经元中HIT化合物的功能效率。这些小分子最终将为我们的团体提供一个平台，以后的R01规模提案，以启动全尺度药物发现运动；但是在此提案中，热门化合物将作为要进一步确定（通过NMR）哪种特定侧链与氧化方法驱动的问题 αSyn的错误折叠。呢