Date of Award
Doctor of Philosophy
Equilibrium constants for the binding of concanavalin A (Con A) to two sugars were determined chromatographically using the protein immobilized on diol-modified silica. In the "reversed-role" mode, 4-methylumbelliferyl (alpha)-D-mannopyranoside (MUM) was the solute chromatographed and mannose was the inhibitor in the mobile phase for the temperature range of 4(DEGREES) to 30(DEGREES)C. Plots of the reciprocal of the capacity factor versus inhibitor concentration were found to be linear. Binding enthalpies for MUM of -7.0 kcal/mol and for mannose of -6.6 kcal/mole were obtained;Rate constants were determined using the "peak-decay" method for MUM, p-nitrophenyl (alpha)-D-mannopyranoside (PNPM) and ovalbumin (OVA) as solutes on the immobilized Con A support. Dissociation rate constants at 25(DEGREES)C for MUM of 1.8 s('-1), for PNPM of 2.6 s('-1), and for OVA of 7.2 x 10('-3) s('-1) were obtained. Enthalpies of reaction were found to be less than those for the faster reaction taking place in solution. Multiple equilibria are probably the cause for this observation.
Digital Repository @ Iowa State University, http://lib.dr.iastate.edu/
Robert Michael Moore
Moore, Robert Michael, "Determination of thermodynamic and kinetic properties of concanavalin A and various sugars using high-performance affinity chromatography " (1987). Retrospective Theses and Dissertations. 8568.