摘要
It has been reported that inactivation occurs before noticeable conformational changes can be detected during denaturation of creatine kinase and other enzymes. Therefore, Tsou suggested that enzyme active sites may display more conformational flexibility than the enzyme molecules as a whole. In the present investigation, the conformational changes of Penaeus penicillatus acid phosphatase during denaturation in urea solutions were studied by following changes in the intrinsic fluorescence, ultraviolet difference absorption, and circular dichroism spectra. Inactivation of the enzyme in urea solutions was compared with unfolding of the enzyme molecule. The results show that the extent of unfolding in guanidine solutions measured by several different methods closely coincides with each other and that slightly lower concentrations of guanidine are required to bring about inactivation than are required to produce significant conformational changes of the enzyme molecule. At the same concentrations, the inactivation rate constants are markedly faster than the rate constants for unfolding of the enzyme. The above results suggest that the active sites of this enzyme display more conformational flexibility than the enzyme molecule as a whole.
It has been reported that inactivation occurs before noticeable conformational changes can be detected during denaturation of creatine kinase and other enzymes. Therefore, Tsou suggested that enzyme active sites may display more conformational flexibility than the enzyme molecules as a whole. In the present investigation, the conformational changes of Penaeus penicillatus acid phosphatase during denaturation in urea solutions were studied by following changes in the intrinsic fluorescence, ultraviolet difference absorption, and circular dichroism spectra. Inactivation of the enzyme in urea solutions was compared with unfolding of the enzyme molecule. The results show that the extent of unfolding in guanidine solutions measured by several different methods closely coincides with each other and that slightly lower concentrations of guanidine are required to bring about inactivation than are required to produce significant conformational changes of the enzyme molecule. At the same concentrations, the inactivation rate constants are markedly faster than the rate constants for unfolding of the enzyme. The above results suggest that the active sites of this enzyme display more conformational flexibility than the enzyme molecule as a whole.
