Fourier-transform infared spectroscopy, combined with resolution-enhancement techniques including second-derivative spectroscopy, Fourier self-deconvolution and curvefitting technique, was used to investigate the thermally induced unfolding process of anti-HIV-I toxin protein trichosanthin. During heating from 25 degrees C to 85 degrees C, the peak of Amide I shifted to 1618 cm-1 while the secondary structural contents change with the temperature. Upon cooling the protein from 85 degrees C to 25 degrees C, the contour of the Amide I do not change. All these show that the thermal unfolding of trichosanthin is an irreversible intermolecular aggregation process between 25 degrees C and 85 degrees C. The changes of secondary structures with temperature suggest the presence of folding intermediates.