Capillary Electrophoresis and Protein Biophysics

描述(申请人提供)：这项研究集中于基于蛋白质表面疏水性的新技术的开发，用于分离、表征和操纵蛋白质。(这项提案是对之前一项没有资金的提案进行了大量修改。)改进用于分离、表征和操作蛋白质的方法在生物化学和分子生物学中具有广泛的重要性：应用领域包括蛋白质组学和系统生物学、蛋白质疗法的纯化、用于临床分析的芯片实验室分析系统、合理的药物设计和基础蛋白质科学。建议的研究有五个具体目标：1.开发一种新的、有效的模型系统来研究蛋白质表面疏水性区域的生物物理。2.根据蛋白质对疏水表面活性剂和亲水性物质(如十二烷基硫酸钠，而不是尿素)变性的敏感性，开发表征蛋白质的新技术。3.使使用疏水表面活性剂处理蛋白质的体系合理化。4.解决塑料LOC系统中蛋白质在微通道疏水壁上的吸附问题。5.了解介质的疏水性和有机特性对配体与蛋白质结合的影响。 这项研究将有助于分析和提纯用于治疗人类疾病的蛋白质，并有助于开发在疾病诊断、公共卫生筛查和应对生物事件(新出现的疾病、生物恐怖主义)中有用的仪器技术。

DESCRIPTION (provided by applicant): This research is focused on the development of new techniques, based on the hydrophobicity of their surfaces, for separating, characterizing, and manipulating proteins. (The proposal is an extensively revised version of a previous proposal that was not funded.) Improving the methods used to separate, characterize, and manipulate proteins is broadly important in biochemistry and molecular biology: areas of application include proteomics and systems biology, purification of protein therapeutics, use in lab-on-a-chip analytical systems for clinical analyses, rational drug design, and fundamental protein science. The proposed research has five specific objectives: 1. To develop a new, validated model system for studying the biophysics of hydrophobic regions of protein surfaces. 2. To develop new techniques for characterization of proteins based on their susceptibility to denaturation by hydrophobic surfactants and hydrotropes (e.g., SDS, not urea). 3. To rationalize systems that use hydrophobic surfactants in manipulating proteins. 4. To provide a solution to the problem of adsorption of proteins on the hydrophobic walls of microchannels in plastic LoC systems. 5. To help to understand the influence of hydrophobicity and organic character of the medium on binding of ligands to proteins. Lay Description: This research will contribute to the analysis and purification of proteins used in treatment of human disease, and to the development of instrumental techniques useful in the diagnosis of disease, in screening for public health, and in response to biological events (emerging disease, bioterrorism).