Mechanism of protein salting in and salting out by divalent cation salts: balance between hydration and salt binding.
The preferential interactions of proteins with solvent components were studied in concentrated aqueous solutions of the sulfate, acetate, and chloride salts of Mg2+, Ba2+, Ca2+, Mn2+ and Ni2+ [except for CaSO4, BaSO4, Mn-(OAc)2, and Ni(OAc)2], and results were compared with those of the Na+ salts. It was found that, for all the salts, the preferential hydration increased in the order of Cl- less than CH3-COO- less than SO42- regardless of the cationic species used, in agreement with the anionic lyotropic series, and that the same parameter exhibited a tendency to increase in the order of Mn2+, Ni2+ less than Ca2+, Ba2+ less than Mg2+ less than Na+. The salting-out and stabilizing or salting-in and destabilizing effectiveness of the salts were interpreted in terms of the observed preferential interactions. The surface tension increment of salts, which is a major factor responsible for the preferential interactions of the Na+ salts, had no correlation with those of the divalent cation salts. It was shown that the binding of divalent cations to the proteins overcomes the salt exclusion due to the surface tension increase, leading to a decrease in the preferential hydration. In conformity with this mechanism, the preferential interaction of MgCl2 was strongly pH dependent, because of the protein charge-dependent affinity of Mg2+ for proteins, while NaCl showed no pH dependence of the preferential interaction. The proposed mechanism was supported by a strong correlation between the preferential interaction results and the interaction of these salts with the model peptide compound acetyltetraglycine ethyl ester, described by Robinson and Jencks.
D000085:Acetates; D001464:Barium / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002712:Chlorides; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D009532:Nickel / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D012492:Salts / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D012995:Solubility; D012997:Solvents; D013431:Sulfates; D013816:Thermodynamics
D000085:Acetates; D002413:Cations, Divalent; D002712:Chlorides; D011506:Proteins; D012492:Salts; D012997:Solvents; D013431:Sulfates; D001464:Barium; D012710:Serum Albumin, Bovine; D008345:Manganese; D009532:Nickel; D009113:Muramidase; D008274:Magnesium; D002118:Calcium