Journal of Inherited Metabolic Disease

, Volume 19, Issue 3, pp 278–285 | Cite as

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

  • E. A. Karpova
  • Ya. V. Voznyi
  • J. L. M. Keulemans
  • A. T. Hoogeveen
  • B. Winchester
  • I. V. Tsvetkova
  • O. P. van Diggelen
Article

Summary

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.

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References

  1. Freeman C, Hopwood JJ (1986) Human liver sulphamate sulphohydrolase.Biochem J 234: 83–92.Google Scholar
  2. Hopwood JJ, Elliott H (1981a) Sulphamidase activity in leucocytes, cultured skin fibroblasts and amniotic cells: diagnosis of Sanfilippo A syndrome with the use of a radiolabelled disaccharide substrate.Clin Sci 61: 729–735.Google Scholar
  3. Hopwood JJ, Elliott H (1981b) Radiolabelled oligosaccharides as substrates for the estimation of sulfamidase and the detection of Sanfilippo type A syndrome.Clin Chim Acta 112: 55–66.Google Scholar
  4. Hopwood JJ, Elliott H (1982) Diagnosis of Sanfilippo type A syndrome by estimation of sulfamidase activity using a radiolabelled tetrasaccharide substrate.Clin Chim Acta 123: 241–250.Google Scholar
  5. Kresse H (1973) Mucopolysaccharidosis IIIA (Sanfilippo A disease): deficiency of heparan sulphamidase in skin fibroblasts and leucocytes.Biochem Biophys Res Commun 54: 1111–1118.Google Scholar
  6. Neufeld EF, Muenzer J (1995) The mucopolysaccharidoses. In Scriver CH, Beaudet AL, Sly WS, Valle D, eds.The Metabolic and Molecular Bases of Inherited Disease. New York: McGraw-Hill, 2465–2495.Google Scholar
  7. Van Diggelen OP, Zhao H, Kleijer WJ, et al (1990) A fluorimetric enzyme assay for the diagnosis of Morquio disease type A (MPS IVA).Clin Chim Acta 187: 131–140.Google Scholar
  8. Voznyi Ya V, Afanasyeva SV, Kalicheva IS, Galoyan AA (1991) 2-Deoxy-2-trifluoroacetamido-β-D-glucopyranosyl fluoride in the synthesis of fluorogenicα- andβ-N-acetylglucosaminides.Bioorganicheskaya Khimia 17: 1251–1257.Google Scholar
  9. Voznyi Ya V, Karpova EA, Dudukina TV, et al (1993) A fluorimetric enzyme assay for the diagnosis of Sanfilippo disease C (MPS IIIC).J Inher Metab Dis 16: 465–472.Google Scholar
  10. Whiteman P, Young E (1977) The laboratory diagnosis of Sanfilippo disease.Clin Chim Acta 76: 139–147.Google Scholar

Copyright information

© SSIEM and Kluwer Academic Publishers 1996

Authors and Affiliations

  • E. A. Karpova
    • 4
  • Ya. V. Voznyi
    • 1
  • J. L. M. Keulemans
    • 4
  • A. T. Hoogeveen
    • 4
  • B. Winchester
    • 2
  • I. V. Tsvetkova
    • 3
  • O. P. van Diggelen
    • 4
  1. 1.Institute of Organic ChemistryMoscowRussia
  2. 2.Division of Biochemistry and GeneticsInstitute of Child HealthLondonUK
  3. 3.Institute of Biomedical ChemistryMoscowRussia
  4. 4.Department of Clinical GeneticsErasmus UniversityRotterdamThe Netherlands

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