An enzymatic method for the assay of serum argininosuccinate lyase.
A rapid enzymatic method was developed for the assay of serum argininosuccinate lyase (ASAL: EC 4.3.2.1) which is a useful marker enzyme for diagnosis of parenchymal liver diseases. Fumarate, liberated from argininosuccinate in the lyase-mediated reaction, was converted to pyruvate via L-malate by the actions of fumarase and malic enzyme in the presence of NADP+. The NADPH formed was then oxidized with a diaphorase-resazurin system to give a highly fluorescent resorufin. All the enzymatic reactions proceeded continuously in 0.1 M Tris-HCl buffer (pH 7.5) and allowed direct assay of ASAL in serum by monitoring the increase in the fluorescence intensity due to resorufin. The method is rapid and sensitive; only 50 microliter of serum is required. This method was used to detect increases in the activities in sera from patients with liver diseases.
Miura
T
T
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Kashiwamura
M
M
Kimura
M
M
D001123:Argininosuccinate Lyase / Q000097:blood*; D008058:Dihydrolipoamide Dehydrogenase; D005649:Fumarate Hydratase; D006801:Humans; D008107:Liver Diseases / Q000201:enzymology; D008190:Lyases / Q000097:blood*; D008291:Malate Dehydrogenase; D009249:NADP; D010078:Oxazines; D013050:Spectrometry, Fluorescence
D010078:Oxazines; D009249:NADP; C014180:resorufin; D008291:Malate Dehydrogenase; D008058:Dihydrolipoamide Dehydrogenase; D008190:Lyases; D005649:Fumarate Hydratase; D001123:Argininosuccinate Lyase