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A sensitive radiometric assay for tryptophan hydroxylase applicable to crude extracts
Abstract:
We describe here a simple and convenient method for assay of tryptophan 5-monooxygenase (hydroxylase), applicable to enzyme in all states of purification. It is based on the enzyme-catalysed formation of 5-hydroxy-[4-3H]tryptophan from [5-3H]tryptophan, and the subsequent acid-dependent quantitative release of 3H as 3H2O; unreacted substrate is removed with activated charcoal. The assay is linear with respect to both protein concentration and time, and gives results similar to those in a standard fluorimetric assay.
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