Denaturation of adenosine deaminase with urea and guanidine hydrochloride

The denaturation effect of urea and guanidine hydrochloride on the adenosine deaminase has been investigated spectrophotometrically at the two temperatures of 27 degrees C and 37 degrees C at pH = 7.50, phosphate buffer (55 mM). A simple, reversible two state transition, N double left right arrow D, was used to analyze the denaturation process from which conformational stability was estimated using three different methods, namely, the linear extrapolation method (LEM), Tanford's model (TM), and the denaturant binding method (DBM). A good agreement was observed among these methods. The results from free energy of denaturation at zero concentration of denaturant, Delta G(H2O)degrees, show the fragile conformation for adenosine deaminase molecule.