The effect of temperature on gluten conditioned at the following water contents, 0%, 13%, and 47% (wet weight basis), was studied by FTIR spectroscopy over the temperature range of 25-85 degrees C. A detailed discussion of the assignment of the amide I band is given. At 0% hydration no changes in the secondary structure with temperature could be detected; spectra were consistent with a tight disordered structure with many protein-protein interactions. At 13% hydration, distinctive changes occurred in the low-frequency region of the amide I band (1630-1613 cm(-1)). This was attributed to changes in the beta-sheet structure. On cooling to 25 degrees C, these changes were mainly reversed. It was noted that most of the changes observed occurred above the glass transition temperature. At 47% hydration, more complex changes took place: as the temperature was raised distinct bands at 1630 and 1613 cm(-1) merged. However, this process was partially reversed, with recovery of both bands, on cooling. The significance of these results in relation to other changes in gluten proteins in flour and dough with temperature and water content is discussed.