Molecular Basis of the Effects of Hydrostatic Pressure on Protein Stability

静水压力对蛋白质稳定性影响的分子基础

• 批准号: 1145407
• 负责人: George Makhatadze
• 金额: $ 25万
• 依托单位: Rensselaer Polytechnic Institute
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1145407&HistoricalAwards=false
• 关键词: Molecular; Basis; Effects; Hydrostatic; Pressure

In this award from the Chemistry of Life Processes Program in the Chemistry Division, Dr. George I. Makhatadze, from the Rensselaer Polytechnic Institute, will investigate the effects of hydrostatic pressure on the stability of proteins and enzymes. This work is the first step in understanding quantitatively the relative contributions of changes in volumetric properties (volume and thermal expansion coefficient) on protein unfolding. Site-directed variants of model proteins (eglin c, ubiquitin, acylphosphatase) with large-to-small non-polar substitutions in the protein core will be studied using the novel method of pressure perturbation calorimetry, which allows direct measurements of volume changes upon unfolding. The results of these experiments will be combined with structural and computational methods to provide quantitative estimates of the contribution of cavity formation and non-polar hydration to volume changes upon protein unfolding.The proposed experiments will provide the first systematic study of the detailed mechanism of the effects of hydrostatic pressure on protein stability using a combination of site-directed mutagenesis and a variety of biophysical and computational tools. It is expected that the knowledge accumulated as a result of these experiments will lay the foundation for future studies of these fundamentally important issues, with broad implications for the use of proteins and enzymes in biotechnology, including renewable energy, green catalysts, and new biomaterials. Dr. Makhatadze is actively involved in graduate and undergraduate education and curriculum development. In addition, students at all levels (high-school, undergraduate and graduate) are participating directly in both the computational and experimental aspects of all research efforts in the his laboratory.

伦斯勒理工学院的George I.Makhatadze博士将研究静水压力对蛋白质和酶稳定性的影响，该奖项由化学部生命过程化学计划颁发。这项工作是定量了解体积性质(体积和热膨胀系数)变化对蛋白质展开的相对贡献的第一步。模型蛋白质(Eglin c、泛素、酰基磷酸酶)在蛋白质核心具有从大到小的非极性取代的定点变体将被用新的压力微扰量热法研究，该方法允许直接测量展开时的体积变化。这些实验的结果将与结构和计算方法相结合，提供空洞形成和非极性水化对蛋白质展开时体积变化的贡献的定量估计。拟议的实验将结合定点突变和各种生物物理和计算工具，首次系统地研究静水压力对蛋白质稳定性影响的详细机制。预计这些实验积累的知识将为未来对这些根本重要问题的研究奠定基础，并对蛋白质和酶在生物技术中的使用产生广泛影响，包括可再生能源、绿色催化剂和新的生物材料。Makhatadze博士积极参与研究生和本科生教育以及课程开发。此外，所有级别的学生(高中生、本科生和研究生)都直接参与HIS实验室所有研究工作的计算和实验方面。