NIH DIRECTOR'S PIONEER AWARD

美国国立卫生研究院院长先锋奖

• 批准号: 7292761
• 负责人: LILA M GIERASCH
• 金额: $ 78.5万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 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/7292761
• 关键词: Adopted; Amino Acid Sequence; Anabolism; Award; Bacterial Proteins; Cells; Collaborations; Complex; Condition; Defect; Disease; Eukaryotic Cell; Fluorescence; In Vitro; Label; Lead; Methods; Modeling; Nuclear Magnetic Resonance; Organism; Pattern; Process; Proteins; Reaction; Reliance; Reporting; Research; Signal Transduction; Structure; System; Testing; Therapeutic Intervention; Tube; cellular imaging; fitness; in vivo; interest; novel; protein expression; protein folding; protein function; Adopted; Amino Acid Sequence; Anabolism; Award; Bacterial Proteins; Cells; Collaborations; Complex; Condition; Defect; Disease; Eukaryotic Cell; Fluorescence; In Vitro; Label; Lead; Methods; Modeling; Nuclear Magnetic Resonance; Organism; Pattern; Process; Proteins; Reaction; Reliance; Reporting; Research; Signal Transduction; Structure; System; Testing; Therapeutic Intervention; Tube; cellular imaging; fitness; in vivo; interest; novel; protein expression; protein folding; protein function

Surprisingly little is known about how proteins fold in vivo, yet it is this process, and not the test-tube idealized folding reaction so intensively studied over the past several decades, that is crucial to the fitness of an organism. The fidelity of folding and the stability of proteins in the cell are critical to their functions, their degradation, and their vulnerability to aggregation. Many diseases are now known to arise from defects in protein folding, either because of the loss or alteration of essential protein functions, or because of the build-up of toxic species such as aggregates. We believe that novel methods and creative collaborations will allow us to overcome the daunting technical obstacles that have impeded progress on protein folding in the cell. Focusing first on a small model protein for which we have detailed descriptions of folding in vitro will enable methods optimization. Folding in cellular conditions will be followed in systems of increasing complexity: bacterial protein expression, cell-free biosynthesis, and semi-permeabilized or intact eukaryotic cell expression. The new strategies will reveal how larger proteins of biomedical interest adopt their structures in their cellular context and how this process may go awry. Methodologically, we anticipate placing major reliance on spectroscopic methods, including fluorescence and nuclear magnetic resonance, and using novel labeling strategies to observe the protein under study in the complex cellular milieu. Complementary in-cell imaging methods will be used to insure that observed signals report on relevant phenomena and to reveal novel functionally significant spatial localization patterns. We anticipate that this research will lead to new paradigms for how amino acid sequences encode folding information and that the resulting enhanced understanding of folding in vivo will lead to new strategies for therapeutic intervention in misfolding and aggregation diseases.

令人惊讶的是，关于蛋白质如何在体内折叠的知之甚少，但这是该过程，而不是试管 在过去的几十年中，理想化的折叠反应如此深入研究，这对 有机体的健身。折叠的保真度和蛋白质在细胞中的稳定性对它们至关重要 功能，降解及其易受聚合的脆弱性。现在已经知道许多疾病 是由于蛋白质折叠的缺陷而引起的，要么是由于必需蛋白质的损失或改变 功能，或者是因为有毒物种（例如聚集体）的积累。我们相信那本小说 方法和创意合作将使我们能够克服艰巨的技术障碍 在细胞中的蛋白质折叠上阻碍了进展。首先专注于小型模型蛋白 我们对体外折叠的详细描述将启用方法优化。在细胞中折叠 在增加复杂性的系统中，将遵循条件：细菌蛋白表达，无细胞 生物合成以及半渗透或完整的真核细胞表达。新策略将 揭示生物医学兴趣的较大蛋白质如何在细胞环境中采用其结构以及如何采用 这个过程可能会出错。从方法上讲，我们预计将对光谱镜的重大依赖 方法，包括荧光和核磁共振，并使用新型的标记策略 观察复杂细胞环境中研究的蛋白质。互补的内部成像 方法将用于确保观察到有关相关现象的信号报告并揭示新颖的信号 功能上重要的空间定位模式。我们预计这项研究将导致新 氨基酸序列如何编码折叠信息的范例以及所得增强的 了解体内折叠的理解将导致新的策略，以进行错误折叠和 聚集疾病。