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

该项目由分析和表面化学项目支持，使用正面分析连续毛细管色谱（FACCE）来表征蛋白质与聚电解质的结合。该项目由印第安纳大学-普渡大学的Paul L. Dubin博士和他的团队进行，该项目将face的范围扩大到包括pi或pi以下的蛋白质。通过在毛细管壁上物理吸附阳离子-非离子嵌段共聚物来修饰FACCE中使用的毛细管，从而扩展了该范围。结合等温线和结合常数（Kb）得到了多阴离子蛋白对的生物学兴趣。特别强调的是糖胺聚糖，特别是透明质酸和肝素。face研究还将对肝素和血清白蛋白等偶对进行研究，以阐明聚合物电荷密度和链刚度等整体大分子结构参数的作用。蛋白质静电的计算建模有助于合理的影响关键变量，如离子强度和pH值对Kb值通过face获得。本研究使用正面分析连续毛细管色谱（FACCE）和蛋白质静电来表征蛋白质与聚电解质的结合。所开发的信息将增加对这种结合在活生物体的细胞间信号传导和细胞发育中的作用的理解。

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.