An enzymatic method for the assay of serum argininosuccinate lyase.

Abstract

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.

• Authors: T Miura, M Kashiwamura, M Kimura
• Journal: Analytical biochemistry (Anal Biochem) Vol. 164 Issue 2 Pg. 482-7 (Aug 01 1987) ISSN: 0003-2697 [Print] United States
• PMID: 3674395 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Oxazines
• NADP
• resorufin
• Malate Dehydrogenase
• Dihydrolipoamide Dehydrogenase
• Lyases
• Fumarate Hydratase
• Argininosuccinate Lyase

Topics

• Argininosuccinate Lyase (blood)
• Dihydrolipoamide Dehydrogenase
• Fumarate Hydratase
• Humans
• Liver Diseases (enzymology)
• Lyases (blood)
• Malate Dehydrogenase
• NADP
• Oxazines
• Spectrometry, Fluorescence