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A Fluorometric Assay for Glycosylasparaginase Activity and Detection of Aspartylglycosaminuria

A Fluorometric Assay for Glycosylasparaginase Activity and Detection of Aspartylglycosaminuria

I T Mononen, V M Kaartinen, J C Williams

Anal Biochem. 1993 Feb 1;208(2):372-4.

PMID: 8452235

Abstract:

Recent experimental work on the mechanism of action of glycosylasparaginase suggests that the enzyme specifically reacts toward the L-asparagine or L-aspartic acid moiety of its substrates. Based on this, a new sensitive assay for glycosylasparaginase activity has been developed using L-aspartic acid beta-(7-amido-4-methylcoumarin) as substrate. Release of 7-amino-4-methylcoumarin was determined fluorometrically. At pH 7.5, Km = 93 microM, and as little as 1 ng of glycosylasparaginase could be detected with the assay. Hydrolysis of the substrate was inhibited by diazo-oxonorvaline, a specific inhibitor of glycosylasparaginase. In biological samples, the fluorometric assay is 40-100 times more sensitive than other published methods for glycosylasparaginase. This new assay enables a rapid enzymatic diagnosis of aspartylglycosaminuria--a genetic deficiency of glycosylasparaginase activity--with leukocyte and fibroblast samples.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP133628736	L-Aspartic acid β-(7-amido-4-methylcoumarin)		133628-73-6	Price

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