This document introduces how to use fluorescein to detect the effects of trial drugs on the activity of trypanothione reductase.
The document details a scientific study investigating the photoreactivity of fluphenazine as a photoaffinity label for trypanothione reductase (TR). The research focuses on how phenothiazine inhibitors bind to TR, which is relevant for developing drugs against African sleeping sickness and other parasitic diseases.
The text explains that the Cary 1E Spectrophotometer was utilized to follow the photolysis of fluphenazine in various media and to measure changes in the UV-visible spectrum of trypanothione reductase after photoinhibition. The study confirms that fluphenazine acts as a competitive inhibitor and undergoes irreversible, time-dependent inactivation when photolyzed in the presence of TR.
The study utilized a Cary 1E spectrophotometer equipped with a Cary temperature controller and an Applied Photophysics Model 2007 Quantum Yield Photoreactor.
The solutions contained 0.03 mM fluphenazine in dimethyl sulphoxide and 0.02 mM in Hepes buffer.
The samples were irradiated using a source of incident light with a wavelength range above 300 nm.