Structural studies of arginine induced enhancement in the activity of T7 RNA polymerase

Abstract

Addition of arginine enhances the activity of the enzyme T7 RNA polymerase. Different methods have been employed to understand the enhancement in the light of arginine induced alteration of the tertiary structure. The increase in activity of the enzyme reaches a maximum value around a concentration of 125mM arginine. Fluorescence, circular dichroism and dynamic light scattering studies indicate an alteration in the tertiary structure of the enzyme. Enthalpy change as a function of input concentration of arginine to a fixed concentration of the enzyme (5μM) shows a dip at 100mM concentration of arginine. Differential scanning calorimetric studies of the denaturation of the enzyme in absence and presence of arginine indicates arginine induced destabilization of the C-terminal domain of the enzyme. Structural alterations induced by arginine have been compared with those induced by the denaturant guanidine hydrochloride.