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。聚集过程，没有成功的化合物在PD的分子基础上进行研究。最近已经发生了巨大的转变，专注于有毒寡聚αSyn，与神经变性，理解这一有希望的新治疗靶原纤维，现在需要生物物理洞察αSyn单体折叠到有毒的寡聚形式中蛋白质的几项研究表明，氧化应激条件是形成的剧毒αSynoligomers。氧化的αSyn。 C末端酪氨酸残基在错误折叠过程中的重要性。这是基于自己的最新发现（2016年在自然化学生物学上发表）蛋氨酸的of导致形成强烈的非共价相互作用，包括酪氨酸。我们的方法将提供化学和生物物理学的定量细节 NMR的测量和计算建模。分子在forst时期靶向有毒，氧化的αsynoligomers的形成和稳定性将利用一个强大的新平台进行高通量筛选，该平台基于精致敏感的荧光寿命测量值，并测试神经元中的HIT化合物的功能。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，以启动全尺度药物药物发现运动；探针进一步确定（通过NMR）哪种特定的侧链相互作用与氧化的甲氧氨酸驱动 αSyn的错误折叠。呢