The influence of some methodological factors on measurement of tryptophan oxygenase activities in crude homogenates of rat liver

Abstract

With two different methods for assaying the tryptophan oxygenase activity in rat liver homogenates, the effects of some methodological factors on the activity of the enzyme were studied. In fed, but not in starved, rats (a) compound(s) absorbing at 365 nm, interfering with the reading of kynurenine absorbance, disappeared gradually during incubation. A correction for this tryptophan-independent reaction was necessary in order to determine correct tryptophan oxygenase activity. Blood remaining in liver tissue post mortem can serve as a source of cofactor haem for tryptophan oxygenase, causing spuriously high values for the activity of the holoenzyme form of tryptophan oxygenase. A rapid and progressive activation of tryptophan oxygenase post mortem occurs in undisrupted liver tissue, and this activation is temperature-dependent.