Temperature dependence of binding of hyaluronate oligomer to bovine nasal proteoglycan

Abstract

An ultrafiltration technique was used to study the temperature coefficient of the association constant K for 1:1 binding of proteoglycan to a hyaluronate oligosaccharide fraction containing an average of about 16 monosaccharide units. The proteoglycan was concentrated during the filtration experiment in order to provide minimal disturbance of the equilibrium in the retained solution. Analytical results calculated from assay of 3H-labeled hyaluronate in the filtrate fractions were extrapolated back to initial equilibrium cell conditions. At 10 °C values of K obtained in this way from ultrafiltration agreed within experimental error with those from equilibrium dialysis. Apparent K values obtained with both techniques tended to decrease somewhat with increasing proteoglycan concentration, due probably in part to excluded volume effects. Values of K obtained by ultrafiltration over the temperature range 5 to 40 °C were used to estimate the enthalpy of binding Δ H∘ as −17.5 (±1.5) kcal mol −1 and the entropy of binding Δ S∘ as −50 (±5) cal K −1 mol −1 (based on a 1 μ m standard state). The dilute solution value of K at 37 °C is sufficiently large to suggest that most of the proteoglycan monomers having a binding site are complexed under tissue conditions.