Melting-profile analysis of thermal stability of thermolysin. A formulation of temperature-scanning kinetics

Abstract

The melting-profile method consists of a continuous observation of a structural parameter while the temperature of the sample is raised at a constant rate. An analytical solution to the melting profile was formulated from the 2-state irreversible process and called temperature-scanning kinetics. The theory was tested with thermolysin [EC 3.4.24.4] with consistent results, and the thermodynamic parameters of thermal denaturation were calculated: .**GRAPHIC**. = 80.3 kcal/mol, .**GRAPHIC**. = 153 eu [entropy units]. These values agreed with the corresponding values obtained from the classical constant-temperature relaxation kinetics. The possibilities of temperature-scanning kinetics are discussed.