Glycosylation of hemoglobin S by reducing sugars and its effect on gelation.

The binding of various reducing mono- and disaccharides to hemoglobin S has been measured both before and after treatment of the sugar-protein adducts with NaBH4. Incubation of 0.3 M solutions of D-glucose, D-galactose, D-maltose, and lactose, with 2% hemoglobin for 2 h at 37 degrees C, pH 7.2, leads to the incorporation of 1.1, 1.8, 1.8, and 3.3 mol of sugar, respectively, into 1 mol of hemoglobin tetramer (either A or S). Exposure of these aldose-protein adducts to NaBH4 for an additional hour at 10 degrees C increases the binding to 2.0, 3.3, 2.5, and 4.1 mol per mol tetramer, as would be expected if Schiff base linkages were involved in this protein modification reaction. The data suggest a stereochemical requirement for enhanced binding. The dependence of the pre-reduction binding of glucose on the sugar concentration, and on the oxygenation state of hemoglobin has also been examined. Glycosylation of hemoglobin significantly increases the minimum gelling concentration of the deoxy conformation, as measured by sedimentation equilibrium ultracentrifugation. Of the sugar derivatives of hemoglobin S examined by this method, those modified by D-galactose or lactose have minimum gelling concentrations (in the absence of 2,3-diphosphoglycerate) which are comparable to, or greater than, that of fully carbamylated hemoglobinS.