Un nouveau colorant fluorescent pour la mesure du temps de relaxation brownienne des proteines

Publisher Website
Google Scholar
Abstract
Several authors have described the NBD-chloride (7-chloro-4-nitrobenzo-2-oxa-1,3-diazole) as a fluorogenic reagent of the amino groups and the sulfhydryl groups. It reacts rapidly with the trypsin to produce a stable and highly fluorescent conjugate. The determination of the labelled residues was made with thin layer chromatographies; the lysin residues were preferentially labelled. Fluorescence depolarization measurements of the conjugate were used to determine the brownian relaxation of the enzyme. The temperature dependence of the life-time is very strong and the corrected Perrin's plot demonstrates that the relaxation time of the enzyme decreases rapidly with the temperature. The conclusion is that the NBD-chloride can be used, like the DNS-chloride but is more reactive and stable and its fixation, on trypsin can be followed by measuring the absorbance of the dye at 470 nm.