NIH Director's Pioneer Award

NIH 院长先锋奖

• 批准号: 7292760
• 负责人: GARY JOSEPH PIELAK
• 金额: $ 73万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-28 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7292760
• 关键词: Award; Behavior; Biochemical; Biochemical Process; Biological; Biology; Cells; Condition; Crowding; Data; Disease; Escherichia coli; Eukaryotic Cell; Goals; In Vitro; Knowledge; Life; Measures; Molecular; NMR Spectroscopy; Neurodegenerative Disorders; Physiological; Play; Property; Protein Chemistry; Proteins; Range; Role; Structure; Techniques; Testing; Tube; United States National Institutes of Health; Work; base; enzyme mechanism; frontier; human disease; in vivo; polypeptide; protein protein interaction; protein structure function; synuclein; tau Proteins; theories

My long term goal is to understand the effects of macromolecular crowding on biochemical processes by acquiring atomic-level information on proteins under actual biological conditions. To this end, my group has pioneered in-cell NMR. We have shown how to assess structure, quantify dynamics, and measure stability under the crowded conditions found in living Escherichia coli cells using this powerful new technique. Now, we want to take the next step. With support from a Pioneer Award, we will focus on using in-cell NMR in eukaryotic cells to study two key proteins in neurodegenerative diseases, the intrinsically disordered proteins, a-synuclein and tau. These proteins are excellent candidates not only because of their disease relevance, but also because we know that macromolecular crowding has extremely large effects on the properties of disordered proteins. Our understanding of protein structure and function has grown enormously in the last 100 years. We have progressed from pondering what role, if any, polypeptides play in the cell, to unraveling, at the atomic level, the mechanisms of enzymes and the molecular bases of protein-protein interactions vital to understanding human disease. Our accumulating wealth of knowledge has largely come from in vitro studies performed under conditions far different from those found in biology. For example, most biochemical examinations of protein behavior are performed at concentrations in the ?g-to-mg-per-mL range, but the insides of cells, where most proteins perform their work, have protein concentrations of >300 mg per mL. Thus, our knowledge comes from data acquired under conditions that are far from physiological relevant, and theory predicts these differences can have extremely large effects on biophysical parameters. Moving beyond the test tube by performing truly in vivo studies in living eukaryotic cells by using NMR spectroscopy is the next frontier in protein chemistry.

我的长期目标是了解大分子拥挤的影响 通过获取有关蛋白质的原子级信息的生化过程 实际生物条件。为此，我的小组已经开创了核心内NMR。 我们已经展示了如何评估结构，量化动力学和测量 在活着的大肠杆菌细胞中发现的拥挤条件下的稳定性 这种强大的新技术。现在，我们想迈出下一步。 在先驱奖的支持下，我们将专注于使用内部NMR 真核细胞研究神经退行性疾病中的两个关键蛋白， 本质上无序的蛋白质，A-核蛋白和tau。这些蛋白质是 优秀的候选人不仅是因为他们的疾病相关，还因为 因为我们知道大分子拥挤对 无序蛋白质的特性。 我们对蛋白质结构和功能的理解在 最近100年。我们已经从思考角色（如果有的话）方面发展 多肽在细胞中发挥作用，在原子水平上揭示机制 酶的酶和蛋白质蛋白相互作用的分子碱基对 了解人类疾病。 我们积累的丰富知识主要来自体外研究 在与生物学中发现的条件不同的条件下进行的。为了 例如，大多数蛋白质行为的生化检查是在 每毫升范围内的浓度浓度，但是细胞的内部，其中大多数 蛋白质执行其工作，蛋白质浓度为每毫升> 300 mg。 因此，我们的知识来自在条件下获取的数据 从生理相关性，理论可以预测这些差异可以 对生物物理参数的极大影响。超越试管 通过使用NMR在活体真核细胞中进行真正的体内研究 光谱法是蛋白质化学的下一个领域。