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Detection of the Sanfilippo D syndrome by the use of a radiolabeled monosaccharide sulfate as the substrate for the estimation of N-acetylglucosamine-6-sulfate sulfatase
- Source :
- Analytical Biochemistry. 138:205-209
- Publication Year :
- 1984
- Publisher :
- Elsevier BV, 1984.
-
Abstract
- N -Acetylglucosamine-6-sulfate sulfatase activity was assayed by incubation of the radiolabeled monosaccharide N -acetylglucosamine [1- 14 C]6-sulfate (GlcNAc6S) with homogenates of leukocytes and cultured skin fibroblasts and concentrates of urine derived from normal individuals, patients affected with N -acetylglucosamine-6-sulfate sulfatase deficiency (Sanfilippo D syndrome, mucopolysaccharidosis type IIID), and patients affected with other mucopolysaccharidoses. The assay clearly distinguished affected homozygotes from normal controls and other mucopolysaccharidosis types. The level of enzymatic activity toward GlcNAc6S was compared with that toward a sulfated disaccharide and a sulfated trisaccharide prepared from heparin. The disaccharide was desulfated at the same rate as the monosaccharide and the trisaccharide at 30 times that of the monosaccharide. Sulfatase activity toward glucose 6-sulfate and N -acetylmannosamine 6-sulfate was not detected. Sulfatase activity in fibroblast homogenates with GlcNAc6S exhibited a pH optimum at pH 6.5, an apparent K m of 330 μmol/liter, and inhibition by both sulfate and phosphate ions. The use of radiolabeled GlcNAc6S substrate for the assay of N -acetylglucosamine-6-sulfate sulfatase in leukocytes and skin fibroblasts for the routine enzymatic detection of the Sanfilippo D syndrome is recommended.
- Subjects :
- Mucopolysaccharidosis
Biophysics
Disaccharide
Disaccharides
Biochemistry
Substrate Specificity
Mucopolysaccharidosis III
chemistry.chemical_compound
Sulfation
Leukocytes
medicine
Humans
Monosaccharide
Trisaccharide
Molecular Biology
Cells, Cultured
Skin
chemistry.chemical_classification
Mucopolysaccharidosis Type IIID
Sulfatase
Monosaccharides
Cell Biology
Clinical Enzyme Tests
Fibroblasts
Mucopolysaccharidoses
medicine.disease
Acetylglucosamine
chemistry
Sulfatases
Trisaccharides
Subjects
Details
- ISSN :
- 00032697
- Volume :
- 138
- Database :
- OpenAIRE
- Journal :
- Analytical Biochemistry
- Accession number :
- edsair.doi.dedup.....c086043d6b1732836ec521cb469ee48a