Equilibrium constants and free energies in unfolding of proteins in urea solutions.
A novel thermodynamic approach to the reversible unfolding of proteins in aqueous urea solutions has been developed based on the premise that urea ligands are bound cooperatively to the macromolecule. When successive stoichiometric binding constants have values larger than expected from statistical effects, an equation for moles of bound urea can be derived that contains imaginary terms. For a very steep unfolding curve, one can then show that the fraction of protein unfolded, B, depends on the square of the urea concentration, U, and is given by [equation: see text] A12 is the binding constant as B-->0, and lambda is a parameter that reflects the augmentation in affinities of protein for urea as the moles bound increases to the saturation number, n. This equation provides an analytic expression that reproduces the unfolding curve with good precision, suggests a simple linear graphical procedure for evaluating A12 and lambda, and leads to the appropriate standard free energy changes. The calculated delta G degree values reflect the coupling of urea binding with unfolding of the protein. Some possible implications of this analysis to protein folding in vivo are described.
Klotz
I M
IM
Department of Chemistry, Northwestern University, Evanston, IL 60208-3113, USA. chemdept@chem.nwu.edu
D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D013816:Thermodynamics; D014508:Urea
D011506:Proteins; D014508:Urea