Effects of thermodynamic nonideality on protein interactions. Equivalence of interpretations based on excluded volume and preferential solvation.
Published results on the stabilization of proteins by sucrose (J.C. Lee and S.N. Timasheff, J. Biol. Chem. 256 (1981) 7193) have been reexamined and interpreted in terms of thermodynamic nonideality. The composition dependence of activity coefficients may be accounted for on a statistical-mechanical basis using the concept of excluded volume. An expression is derived in which the effect of sucrose on determination of the partial specific volume of a protein, previously interpreted in terms of preferential protein solvation, is also seen to be attributable to excluded volume. Gel chromatographic studies of the reversible unfolding of alpha-chymotrypsin are presented which demonstrate temperature- and sucrose-mediated changes in the effective volume of the enzyme. These measurements support the quantitative interpretation of the stabilization in terms of thermodynamic nonideality arising from the difference between covolumes for sucrose and the two isomeric states of alpha-chymotrypsin. By establishing the equivalence of the two approaches that have been used to account for the effects of inert solutes on protein transitions, the present investigation eliminates the need for any distinction between such solutes on the basis of molecular size; and also enhances greatly the potential sensitivity of thermodynamic nonideality as a means of probing protein isomerizations, since greater displacement of the equilibrium position may be effected by small rather than by macromolecular solutes present at the same weight concentrations.
D002918:Chymotrypsin; D002919:Chymotrypsinogen; D007700:Kinetics; D011487:Protein Conformation; D011506:Proteins; D012260:Ribonucleases; D012996:Solutions; D013816:Thermodynamics
D011506:Proteins; D012996:Solutions; D002919:Chymotrypsinogen; D012260:Ribonucleases; D002918:Chymotrypsin