Thermal denaturation of porcine pepsin: a study by circular dichroism.
Thermal denaturation of porcine pepsin in 10% ethanol was studied by circular dichroism (CD) spectroscopy. It was observed that the process is markedly irreversible. The denaturation unfolding process was strongly dependent on the heating rate, as is expected for an unfolding process kinetically controlled due to the presence of an irreversible reaction. Experimentally, we demonstrate the existence of an unfolded (U) state in equilibrium with the native (N) state. The U state is observed to exist at temperatures lower than 45 degrees C. The van't Hoff enthalpy, DeltaH(vH), was determined from direct estimation of the equilibrium constant at several temperatures (DeltaH(vH)=304.3 kJ/mol). To explain the observed behavior, we have considered a Lumry-Eyring model, which takes into account the presence of the U state in addition to N and denatured (D) states (i.e. N<-->U-->D).
Tello-Solís
S R
SR
Area de Biofisicoquímica. Departamento de Química, Universidad Autónoma Metropolitana-Iztapalapa, P.O. Box 55-534, 09340, Iztapalapa, Mexico. srts@xanum.uam.mx
Romero-García
B
B
D000818:Animals; D002942:Circular Dichroism; D000431:Ethanol / Q000737:chemistry; D006358:Hot Temperature; D010434:Pepsin A / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding; D013552:Swine; D013696:Temperature; D013816:Thermodynamics
D000431:Ethanol; D010434:Pepsin A