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

该项目得到了分析和表面化学计划的支持，使用前沿分析连续毛细管色谱(FACE)来表征蛋白质与聚电解质的结合。该项目由印第安纳大学-普渡大学印第安纳波利斯分校的保罗·L·杜宾博士和他的团队进行，该项目将FACE的范围扩大到包括PIS或以下的蛋白质。通过阳离子-非离子嵌段共聚物在毛细管壁上的物理吸附，对FACE中使用的毛细管进行了改性，从而扩大了使用范围。获得了具有生物学意义的聚阴离子-蛋白质对的结合等温线，从而得到了结合常数(Kb)。特别强调糖胺聚糖，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配对进行，以阐明总体大分子结构参数的作用，如聚合物电荷密度和链刚性。蛋白质静电的计算模型有助于合理解释离子强度和pH等关键变量对FACCE获得的Kb值的影响。本研究使用前沿分析、连续毛细管色谱(FACE)和蛋白质静电学来表征蛋白质与聚电解质的结合。开发的信息将增加对这种结合在生物体内细胞间信号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.