Structural investigation of transglutaminase by Fourier transform infrared spectroscopy.

The secondary structure of transglutaminase was investigated by Fourier transform infrared spectroscopy. Spectra of the protein in both H2O and 2H2O were analyzed by deconvolution and second derivative methods in order to observe the overlapping components of the amide-I band. The quantitative analysis of the amide-I-band components was made by a curve-fitting procedure. The protein was studied in the absence and in the presence of 1 mM GTP, 1 mM Ca2+ and 1 mM GTP/1 mM Ca2+. The quantitative analysis of infrared spectra revealed that no remarkable changes in the secondary structure of the enzyme are induced by GTP, Ca2+ or Ca2+/GTP. Major changes, however were observed in the thermal-denaturation behavior of the protein. The protein showed maximum of denaturation at temperatures over 50-55 degrees C in the absence or in the presence of 1 mM Ca2+ and over 55-60 degrees C in the presence of 1 mM GTP or 1 mM Ca2+/1 mM GTP. The results obtained indicate that GTP induces a stabilization of the tertiary structure of the enzyme, even in the presence of 1 mM Ca2+. The thermal denaturation patterns of the protein suggest the occurrence of Ca(2+)-dependent aggregation.