Experimental and Theoretical Studies of Charge-Charge Interactions in Proteins

蛋白质中电荷相互作用的实验和理论研究

• 批准号: 0416746
• 负责人: George Makhatadze
• 金额: --
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0416746&HistoricalAwards=false
• 关键词: Experimental; Theoretical; Studies; Charge; Interactions

Understanding the mechanisms of stabilization of protein structure represents one of the major aspects of the protein folding problem. The contributions of various short-range interactions, such as hydrogen bonding, the hydrophobic effect, and configurational entropy, to protein stability have been well documented. However, it is becoming more and more evident that the long-range electrostatic interactions between charged protein groups are also important contributors to protein structure and stability. The overall goal of this project is to explore the relation between the charge-charge interactions on the protein surface and properties of proteins such as stability, solubility and enzymatic activity. The experimental work will be performed with several model proteins (ubiquitin, CspB, CDC42) that differ in three dimensional structure, size, and functional activity. The amino acid sequence of these proteins will be altered according to the theoretical predictions based on different models for calculation of charge-charge interactions. The consequences of these changes in terms of stability, solubility and enzymatic activity will be measured using a variety of biophysical methods. The fundamental importance of this research is that using a combination of theoretical calculations and experimental measurements the relationship between charge-charge interactions and the protein stability will be explored.This research will provide the foundation for the design of proteins stable at extreme temperatures. This will benefit the industrial use of proteins where thermostability of enzymes is in many cases the largest obstacle. This research will also have broad impact on the research and education infrastructure because training of graduate and undergraduate students is an intrinsic aspect of all of the experimental and theoretical work. The scientific concepts derived from this work will be incorporated into graduate courses that are offered by the PI.

了解蛋白质结构的稳定机制是蛋白质折叠问题的主要方面之一。各种短程相互作用，如氢键，疏水效应和构型熵，对蛋白质稳定性的贡献已经得到很好的证明。然而，越来越明显的是，带电蛋白质基团之间的长程静电相互作用也是蛋白质结构和稳定性的重要贡献者。本项目的总体目标是探索蛋白质表面的电荷-电荷相互作用与蛋白质性质（如稳定性、溶解性和酶活性）之间的关系。实验工作将使用几种模型蛋白（泛素，CspB，CDC 42）进行，这些蛋白在三维结构，大小和功能活性方面不同。这些蛋白质的氨基酸序列将根据基于用于计算电荷-电荷相互作用的不同模型的理论预测而改变。这些变化在稳定性、溶解度和酶活性方面的结果将使用各种生物物理方法来测量。本研究的重要意义在于通过理论计算和实验测量相结合的方法，探索电荷-电荷相互作用与蛋白质稳定性之间的关系，为设计在极端温度下稳定的蛋白质奠定基础。这将有利于蛋白质的工业应用，其中酶的热稳定性在许多情况下是最大的障碍。这项研究也将对研究和教育基础设施产生广泛的影响，因为研究生和本科生的培训是所有实验和理论工作的一个内在方面。从这项工作中得出的科学概念将被纳入PI提供的研究生课程。