Journal of Inherited Metabolic Disease

, Volume 19, Issue 3, pp 278-285

First online:

A fluorimetric enzyme assay for the diagnosis of sanfilippo disease type A (MPS IIIA)

  • E. A. KarpovaAffiliated withDepartment of Clinical Genetics, Erasmus University
  • , Ya. V. VoznyiAffiliated withInstitute of Organic Chemistry
  • , J. L. M. KeulemansAffiliated withDepartment of Clinical Genetics, Erasmus University
  • , A. T. HoogeveenAffiliated withDepartment of Clinical Genetics, Erasmus University
  • , B. WinchesterAffiliated withDivision of Biochemistry and Genetics, Institute of Child Health
  • , I. V. TsvetkovaAffiliated withInstitute of Biomedical Chemistry
  • , O. P. van DiggelenAffiliated withDepartment of Clinical Genetics, Erasmus University

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4-Methylumbelliferyl-α-d-N-sulphoglucosaminide (MU-α-GlcNS) was synthesized and shown to be a substrate for the lysosomal heparin sulphamidase. Sanfilippo A patients' fibroblasts (n=42) and lymphocytes (n=1) showed 0–3% of mean normal heparin sulphamidase activity; in total leukocytes from patients (n=8) sulphamidase activity was clearly deficient. In fibroblasts from obligate heterozygotes for Sanfilippo A, the sulphamidase activity was reduced in 9 out of 10 cases. Heparin sulphamidase desulphates MU-αGlcNS to MU-αGlcNH2 and further hydrolysis during a second incubation is required to liberate 4-methylumbelliferone, which can be measured. Yeastα-glucosidase, which has low but sufficientα-glucosaminidase activity, was used to hydrolyse the reaction intermediate MU-αGlcNH2 to release 4-methylumbelliferone and free glucosamine.