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

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，人们对蛋白质在体内如何折叠知之甚少，但正是这个过程，而不是试管 理想化折叠反应在过去几十年里得到了深入研究，这对于 有机体的适应性。细胞中蛋白质折叠的保真度和稳定性对于其发挥作用至关重要 功能、退化以及聚合的脆弱性。现在已知许多疾病 由蛋白质折叠缺陷引起，要么是由于必需蛋白质的丢失或改变 功能，或由于聚集体等有毒物质的积累。我们相信小说 方法和创造性合作将使我们能够克服令人畏惧的技术障碍 阻碍了细胞中蛋白质折叠的进展。首先关注一个小模型蛋白质 我们对体外折叠进行了详细描述，这将使方法得以优化。折叠在蜂窝中 在日益复杂的系统中将遵循的条件：细菌蛋白表达、无细胞 生物合成，以及半透化或完整的真核细胞表达。新战略将 揭示具有生物医学意义的较大蛋白质如何在细胞环境中采用其结构以及如何 这个过程可能会出错。从方法上讲，我们预计主要依赖光谱法 方法，包括荧光和核磁共振，以及使用新颖的标记策略 在复杂的细胞环境中观察所研究的蛋白质。互补细胞内成像 将使用方法来确保观测到的信号报告相关现象并揭示新的 具有重要功能的空间定位模式。我们预计这项研究将带来新的成果 氨基酸序列如何编码折叠信息的范例以及由此产生的增强 对体内折叠的理解将导致错误折叠和错误折叠治疗干预的新策略 聚集性疾病。