Abnormal solubility behavior of beta-lactoglobulin: salting-in by glycine and NaCl.
The causes of the salting-in of beta-lactoglobulin by glycine and NaCl, a solubility behavior contrary to expectations, were probed by a detailed study of the interactions between these solvent components and the protein. The preferential interactions of beta-lactoglobulin with solvent components in aqueous glycine and NaCl systems have been compared with those of bovine serum albumin and lysozyme. At neutral pH, beta-lactoglobulin exhibited insignificant preferential interactions in glycine and NaCl at low cosolvent concentrations and an increasing preferential hydration at higher concentrations, the levels approaching the values expected from the other two proteins. These results indicate considerable binding of the electrolytes to beta-lactoglobulin, sufficient to compensate for the exclusion due to perturbation of the solvent surface tension. The difference between the preferential interactions of beta-lactoglobulin and the other proteins with these two solvent additives was shown to be the cause of the increase of beta-lactoglobulin solubility even at high concentrations of the additives, at which they have salting-out effects on the other proteins. The preferential interactions of NaCl with the three proteins were examined as a function of pH. The results showed no pH dependence of the preferential hydration for bovine serum albumin and lysozyme, while this parameter increased significantly for beta-lactoglobulin at lower pH. This suggests that the binding of electrolytes to beta-lactoglobulin is due to a unique charge distribution on the surface of the protein around neutral pH, which imparts to this protein a large dipole moment.
Arakawa
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Graduate Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02254.
Timasheff
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D000409:Alanine; D005998:Glycine; D007782:Lactoglobulins; D009113:Muramidase; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D012995:Solubility
D007782:Lactoglobulins; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D009113:Muramidase; D000409:Alanine; D005998:Glycine