Fluorometric coupled-enzyme assay for delta-aminolevulinate synthase

D. F. Bishop, L. McBride, R. J. Desnick

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9 Scopus citations

Abstract

The rapid and specific determination of picomole quantities of δ-aminolevulinate has been accomplished by its enzymatic conversion to uroporphyrinogen I and subsequent fluorometric detection of the oxidized urophorphyrin I. The coupled-enzyme assay was linear with time and protein concentration and required less than 3 h for 25 individual determinations. Under the standard assay conditions, 5-100 pmol of uroporphyrin I was reliably quantitated; these values corresponded to a range of δ-aminolevulinate synthase activities from 0.2 to 15 nmol/h/ml enzyme. The sensitivity of this method was comparable to the more time-consuming radiochemical determinations of δ-aminolevulinate synthase activity. The δ-aminolevulinate synthase activity of liver homogenates from uninduced rats was 8.7 U/g liver (37°C).

Original languageEnglish
Pages (from-to)94-107
Number of pages14
JournalEnzyme
Volume28
Issue number2-3
DOIs
StatePublished - 1982

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