A DIFFERENTIAL SCANNING CALORIMETRIC STUDY OF THE THERMAL UNFOLDING OF 7 MUTANT FORMS OF PHAGE-T4 LYSOZYME

High-sensitivity differential scanning calorimetry has been applied to the study of the reversible thermal unfolding of the lysozyme of T4 bacteriophage in which the threonine residue at position 157 has been replaced by seven different residues. High-resolution structures derived from X-ray crystallography have been reported for these and six other mutants by Alber et al. [Alber, T., Dao-Pin, S., Wilson, K., Wozniak, J. A., Cook, S. P., & Matthews, B. W. (1987) Nature 330, 41-46]. At pH 2.5 the changes relative to the wild-type protein in the standard free energy of unfolding produced by these mutations indicate apparent destabilizations of 0.6 kcal mol-1 (T157R) to 1.9 kcal mol-1 (T157I), whereas the changes in enthalpy of unfolding range from -5.8 kcal mol-1 (T157N) to 11.9 kcal mol-1 (T157E). Since the denaturations are in all cases accompanied by large changes in heat capacity amounting to 2.5 kcal K-1 mol-1, both the free energies and enthalpies are functions of temperature. An intriguing feature of the present results is the relatively large enthalpy changes and the corresponding compensating entropy changes. Our present understanding of the intramolecular energetics of proteins is insufficient to account for these changes.