Quantifying Charge and Stability Influences on Protein Adsorption with Synthetic Mutants of Bacteriophage T4 Lysozyme

量化噬菌体 T4 溶菌酶合成突变体的电荷和稳定性对蛋白质吸附的影响

• 批准号: 9216798
• 负责人: Joseph McGuire
• 金额: $ 20.2万
• 依托单位: Oregon State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-03-01 至 1997-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9216798&HistoricalAwards=false
• 关键词: Quantifying; Charge; Stability; Influences; Protein

This project deals with application of a kinetic model describing the general sequence of events undergone by protein at an interface to quantify protein charge and stability effects on interfacial behavior. Selected synthetic mutants of bacteriophage T4 lysozyme are produced form transformed cells obtained from the Institute of Molecular Biology at the University of Oregon. Mutants exhibiting differences in thermodynamic stability and charge are used to quantify the influences of these molecular properties on reaction rate constants describing protein attachment and unfolding at each of a series of solid surfaces differing in hydrophobicity. Silicon surfaces will be modified by silanization with dichlorodimethylsilane, and in situ ellipsometry will be used to measure the adsorption kinetics and isotherm describing each specific protein- surface contact. Upon completion of the project, current multi-protein adsorption models could be re-examined and refined by studying adsorption from mixtures of lysozyme mutants. Experimentation with more relevant mixtures of proteins would then allow deviations from predicted behavior to be related to properties such as molecular size and shape in a more readily quantifiable way. This approach is expected to result in a sound starting point for quantifying competitive adsorption from complex media and may lead to new separation techniques.

本项目涉及动力学模型的应用 描述蛋白质经历的事件的一般顺序 在一个界面上量化蛋白质电荷和稳定性 对界面行为的影响。 选择的合成突变体 的噬菌体T4溶菌酶是由转化的 从分子生物学研究所获得的细胞， 俄勒冈州大学。 表现出以下差异的突变体 热力学稳定性和电荷被用来量化 这些分子性质对反应速率的影响 描述蛋白质附着和展开的常数 一系列疏水性不同的固体表面。 硅表面将通过硅烷化进行改性， 二氯二甲基硅烷，并在原位椭圆偏振法将使用 测量吸附动力学和等温线描述 每一种特定的蛋白质-表面接触。 完成后 该项目，目前的多蛋白质吸附模型可以 通过研究混合物中的吸附作用， 溶菌酶突变体 更多相关实验 蛋白质的混合物将允许偏离 预测的行为与属性有关， 分子大小和形状更容易量化。 预计这一办法将产生一个良好的起点 用于定量复杂介质中的竞争吸附， 可能会产生新的分离技术。