Studies of the Folding of Thioredoxin by Fragment Complementation on the Development of New Tools to Improve the Understanding of Biomolecular Structure and Function

通过片段互补研究硫氧还蛋白折叠，开发新工具以提高对生物分子结构和功能的理解

• 批准号: 9507255
• 负责人: Maria Luisa Tasayco
• 金额: $ 33万
• 依托单位: CUNY City University of New York
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 1999-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9507255&HistoricalAwards=false
• 关键词: Studies; Folding; Thioredoxin; Fragment; Complementation

9507255 Tasayco The goal of the research is to establish the principles underlying the folding of -sheets. The approach chosen is to study the structure, stability, dynamics and folding of a family of protein fragments. It is proposed to dissect E. coli thioredoxin, a well- characterized small monomeric protein with a mixed -structure, into fragments. They will be generated using tools of biochemistry, chemistry and molecular biology. These fragments will be characterized using a combination of biochemical and biophysical tools with increasing level of structural detail. Circular dichroism in the far and near UV, fluorescence, multidimensional nuclear magnetic resonance spectroscopy and computer modeling will also be used. The educational aspect of this proposal aims to complement the lectures of the standard one-semester Physical Chemistry course for Biochemistry majors using the Inverse Prediction Approach, which consists of building a physical 3D-model of a macromolecular and using it to predict its biophysical properties. %%% The ability to correlate structural changes with ahanges in the biological function holds the key to the understanding of the structure-function relationship of biological macromolecules. However, protein structures are not established at the same pace at which new amino acid sequence are generated. Thus solving the "protein folding problem," that is, predicting the three dimensional structure of a protein based solely on its primary sequence, has become an area of intense research. Much progress has been made in the understanding of the principles behind the formation of one kind of structure commonly found in proteins, the -helix. However, little is known about the formation of -sheets, another equally important structural component. This research will compare the structure, stability and folding of the fragments of thioredoxin and their association products with those of the original protein to elucidate the folding of the still little understood -sheets. ***

9507255 Tasayco该研究的目标是建立基本的原则折叠的纸张。 选择的方法是研究蛋白质片段家族的结构、稳定性、动力学和折叠。 建议解剖E.大肠杆菌硫氧还蛋白是一种特征明确的小单体蛋白， - 结构，变成碎片。 它们将使用生物化学、化学和分子生物学工具生成。 这些片段将使用生物化学和生物物理工具的组合，增加结构细节的水平进行表征。 还将使用远紫外和近紫外圆二色性、荧光、多维核磁共振光谱和计算机建模。该提案的教育方面旨在补充使用反向预测方法的生物化学专业标准一学期物理化学课程的讲座，该方法包括构建大分子的物理3D模型并使用它来预测其生物物理特性。 将结构变化与生物功能变化联系起来的能力是理解生物大分子结构-功能关系的关键。 然而，蛋白质结构的建立与新氨基酸序列的产生并不同步。 因此，解决“蛋白质折叠问题”，即仅根据其一级序列预测蛋白质的三维结构，已成为一个激烈的研究领域。 在理解蛋白质中常见的一种结构形成背后的原理方面已经取得了很大进展， - 螺旋。然而，对另一个同样重要的结构成分-却知之甚少。 本研究将比较硫氧还蛋白片段及其缔合产物与原始蛋白的结构、稳定性和折叠，以阐明仍然知之甚少的-折叠。 ***