Development and Applications of FACCE to the Characterization of Glycosaminoglycan-protein Interactions

FACCE 的开发和应用在糖胺聚糖-蛋白质相互作用表征中的应用

• 批准号: 9987891
• 负责人: Paul Dubin
• 金额: $ 34.94万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9987891&HistoricalAwards=false
• 关键词: Development; Applications; FACCE; Characterization; Glycosaminoglycan

This project, which is supported in the Analytical and Surface Chemistry program uses Frontal Analysis Continuous Capillary Chromatography (FACCE) to characterize the binding of proteins to polyelectrolytes. Conducted by Dr. Paul L. Dubin and his group at Indiana University-Purdue University at Indianapolis, the project extends the range of FACCE to include proteins at or below their pIs. The range is extended by modifying the capillary used in FACCE via physisorption of cationic-nonionic block copolymers on the capillary walls. Binding isotherms and hence binding constants (Kb) are obtained for polyanion-protein pairs of biological interest. Particular emphasis is placed on glycosaminoglycans, especially hyaluronic acid and heparin. FACCE studies will also be carried out with couples such as heparin and serum albumin to elucidate the role of overall macromolecular structural parameters such as polymer charge density and chain stiffness. Computational modeling of protein electrostatics helps rationalize the effects of key variables such as ionic strength and pH on Kb values obtained via FACCE.This research uses Frontal Analysis Continuous Capillary Chromatography (FACCE) and protein electrostatics to characterize the binding of proteins to polyelectrolytes. The information developed will increase the understanding of the role of such binding in intercellular signaling and cell development in living organisms.

该项目得到了分析和表面化学项目的支持，使用前沿分析连续毛细管色谱法（FACCE）来表征蛋白质与聚电解质的结合。 由Paul L博士主持。Dubin和他在印第安纳波利斯的印第安纳州普渡大学的研究小组，该项目扩展了FACCE的范围，包括处于或低于其pI的蛋白质。通过阳离子-非离子嵌段共聚物在毛细管壁上的物理吸附来修改FACCE中使用的毛细管，从而扩展范围。 结合等温线，因此结合常数（Kb）获得的聚阴离子-蛋白质对的生物学利益。 特别强调的是糖胺聚糖，特别是透明质酸和肝素。 FACCE研究也将进行夫妇，如肝素和血清白蛋白，以阐明整体大分子结构参数，如聚合物电荷密度和链刚度的作用。 蛋白质静电的计算模型有助于合理化的关键变量，如离子强度和pH值上的Kb值通过FACCE.This研究获得的影响使用前沿分析连续毛细管色谱（FACCE）和蛋白质静电的特征结合蛋白质的聚电解质。所开发的信息将增加对这种结合在活生物体中的细胞间信号传导和细胞发育中的作用的理解。