Thermodynamic parameters were determined for the thermal denaturation
of Ca2+-bound (holo) and Ca2+-free (apo) alpha-lactalbumin from human
and bovine milk. Thermal denaturation profiles were determined from ch
anges in the intrinsic fluorescence emission intensity (FI) as a funct
ion of temperature(T). Human apo alpha-lactalbumin was heat-denatured
in a 2-state process with T-m = 25 degrees C, Delta H = 167 kJ mol(-1)
, Delta S = 7700 J mol(-1) K-1 and Delta C-p = 15400 J mol(-1) K-1. Th
e corresponding values for bovine apo alpha-lactalbumin were: T-m= 20
degrees C, Delta H = 180 kJ mol(-1),Delta S = 9000 J mol(-1) K-1 and D
elta C-p = 5100 J mol(-1) K-1. Derivative plots of d(FI)/d(T) versus T
revealed that both human and bovine hole cr-lactalbumin were heat-den
atured via a 3-state process. Thermal denaturation transitions were as
sociated with a T-m value of 67 degrees C or 42 degrees C, based on ch
anges in tryptophan or tyrosine FI results, respectively. Apparently C
a2+-bound alpha-lactalbumin possesses two regions (domains) with signi
ficantly different conformational stability. Based on tryptophan fluor
escence measurements, Delta H = 330 kJ mol(-1), Delta S = 4600 J mol(-
1) K-1 and Delta C-p = 8200 J mol(-1) K-1 for human or bovine, hole a-
lactalbumin. From tyrosine fluorescence emission changes, Delta H = 54
-103 kJ mol(-1), Delta S = 300-2000 J mol(-1) K-1 and Delta C-p = 3000
-4000 J mol(-1) K-1.