Differential scanning calorimetry of the irreversible thermal denaturation of thermolysin.
A differential scanning calorimetry study of the thermal denaturation of Bacillus thermoproteolyticus rokko thermolysin was carried out. The calorimetric traces were found to be irreversible and highly scan-rate dependent. The shape of the thermograms, as well as their scan-rate dependence, can be explained by assuming that the thermal denaturation takes place according to the kinetic scheme N k----D, where k is a first-order kinetic constant that changes with temperature, as given by the Arrhenius equation, N the native state, and D the unfolded state or, more probably, a final state, irreversibly arrived at from the unfolded one. On the basis of this model, the value of the rate constant as a function of temperature and the activation energy have been calculated. It is shown that the proposed model may be considered as being one particular case of that proposed by Lumry and Eyring [Lumry, R., & Eyring, H. (1954) J. Phys. Chem. 58, 110] N in equilibrium D----I, where N is the native state, D the unfolded one, and I a final state, irreversibly arrived at from D. Lastly, some comments are made on the use of the scan-rate effect on the calorimetric traces as an equilibrium criterion in differential scanning calorimetry.
Sánchez-Ruiz
J M
JM
Departamento de Química Física, Facultad de Ciencias, Universidad de Granada, Spain.
López-Lacomba
J L
JL
Cortijo
M
M
Mateo
P L
PL
D001407:Bacillus / Q000201:enzymology; D002152:Calorimetry, Differential Scanning; D004795:Enzyme Stability; D006358:Hot Temperature; D008433:Mathematics; D011489:Protein Denaturation; D013816:Thermodynamics; D013820:Thermolysin
D013820:Thermolysin