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

Indiana University, Bloomington IN

Investigators

Abstract

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.