Dehydrogenase denaturation by guanidine hydrochloride measured by fluorescence

Abstract

Denaturation and subsequent renaturation of the enzymes Lactate, Glucose-06-phosphate, Glutamate and Alcohol dehydrogenases, by means of fluorescence spectra and the variation of enzyme activity in each conformational state, have been studied. The denaturating agent has been Guanidine chloride in a range of concentration from 0.5 to 6 M. Special behaviour has been observed in each enzyme in the presence of the denaturating agent. The action of this agent is compared with that of urea. The renaturation percentages obtained are relatively low. Interaction between the denaturating agent and the aminoacids producing the fluorescence of the enzymes is observed.