Journal of Chemical Thermodynamics, Vol.33, No.11, 1511-1525, 2001
Measurement and analysis of results obtained on biological substances with d.s.c.
The differential scanning calorimeter (d.s.c.) has been widely used to determine the thermodynamics of phase transitions and conformational changes in biological systems including proteins, nucleic acid sequences, and lipid assemblies. The d.s.c monitors the temperature difference between two vessels, one containing the biological solution and the other containing a reference solution, as a function of temperature at a given scan rate. Recommendations for d.s.c. measurement procedures, calibration procedures, and procedures for testing the performance of the d.s.c. are described. Analysis of the measurements should include a correction for the time response of the instrument and conversion of the power versus time curve to a heat capacity versus temperature plot. Thermodynamic transition models should only be applied to the analysis of the heat capacity curves if the model-derived transition temperatures and enthalpies are independent of the d.s.c. scan rate. Otherwise, kinetic models should be applied to the analysis of the data. Application of thermodynamic transition models involving two states, two states and dissociation, and three states to the heat capacity versus temperature data are described. To check the operating performance with standard d.s.c.s, samples of (1 to 10) mg (.) cm(3) solutions of hen egg white lysozyme in 0. 1 mol (.) dm(-3) HCl-glycine buffer at pH = (2.4 +/- 0.1) were sent to six different d.s.c. laboratories worldwide. The values obtained from proper measurements and application of a two-state transition model yielded an average unfolding transition temperature for lysozyme of 331.2 K with values ranging between T = 329.4 K and 331.9 K, and an average transition enthalpy of 405 kJ (.) mol(-1) with values ranging from 377 kJ (.) mol(-1) to 439 kJ (.) mol(-1). It is recommended that the reporting of d.s.c. results be specific with regard to the composition of the solution, the operating conditions and calibrations of the d.s.c. determination of base lines that may be model dependent, and the model used in the analysis of the data.
Keywords:differential scanning calorimetry;lipids;phase transition;proteins;transition enthalpy;transition temperature;unfolding transitions