Diagnosis of Maroteaux—Lamy syndrome by the use of radiolabelled oligosaccharides as substrates for the determination of arylsulphatase B activity

Hopwood, John J.; Elliott, Helen; Muller, Vivienne; Saccone, Gino T.P.

doi:10.1042/bj2340507

Cited by 53 publications

(41 citation statements)

References 26 publications

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“…Other enzyme activities Enzyme activity towards sulphamate sulphohydrolase [24], a-L-iduronidase [23], sulphoiduronate sulphohydrolase [25], arylsulphatase B [26], ,-N-acetylhexosaminidase [5], ac-N-acetylglucosaminidase [27], acetyl-CoA: glucosamine N-acetyltransferase [28], ,-glucuronidase [29] and N-acetylgalactosamine-6-sulphate sulphatase [30] substrates were assayed by previously reported procedures.…”

Section: Purification Of 6ssmentioning

confidence: 99%

Human liver N-acetylglucosamine-6-sulphate sulphatase. Purification and characterization

Freeman

Clements

Hopwood

1987

Biochemical Journal

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Human N-acetylglucosamine-6-sulphate sulphatase was purified at least 50000-fold to homogeneity in 780%yield from liver with a simple three-step four-column procedure, which consists of a concanavalin A-Sepharose/Blue A-agarose coupled step, chromatofocusing and Cu2+-chelating Sepharose chromatography. In all, four forms were isolated and partially characterized. Forms A and B, both with a pl greater than 9.5 and representing 30% and 60% respectively of the recovered enzyme activity, were separated by hydroxyapatite chromatography of the enzyme preparation obtained from the Cu2+-chelating Sepharose step. Both forms A and B had native molecular masses of 75 kDa. When analysed by SDS/polyacrylamide-gel electrophoresis, form A consists of a single polypeptide of molecular mass 78 kDa, whereas form B contained 48 kDa and 32 kDa polypeptide subunits. Neither form A nor form B was taken up from the culture medium into cultured human skin fibroblasts. The two other forms (C and D), with pI values of 5.8 and 5.4 respectively, represented approx. 7% and 3% of the total recovered enzyme activity. The native molecular masses of forms C and D were 94 kDa and approx. 75 kDa respectively. Form C contained three polypeptides with molecular masses of 48, 45 and 32 kDa. N-Acetylglucosamine-6-sulphate sulphatase activity was measured with a radiolabelled disaccharide substrate derived from heparin. The development of this substrate enabled the isolation and characterization of N-acetylglucosamine-6-sulphate sulphatase to proceed efficiently. Forms A, B and C had pH optima of 5.0, Km values of 11.7, 14.2 and 11.1 LM respectively and Vmax values of 105, 60 and 53 nmol/min per mg of protein respectively. The molecular basis of the multiple forms of this sulphatase is not known. It is postulated that the differences in structure and properties of the four enzyme forms are due to differences in the state of processing of a large subunit.

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Section: Purification Of 6ssmentioning

confidence: 99%

Human liver N-acetylglucosamine-6-sulphate sulphatase. Purification and characterization

Freeman

Clements

Hopwood

1987

Biochemical Journal

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“…Fibroblast lysates were prepared by seven cycles of freeze-thaw in 0.5 M NaCl, 20 mM Tris-HCl, pH 7. Fibroblast lysates were dialyzed for 16 h against 50 mM sodium formate, pH 3.5, and then specifically assayed for 4S by incubation with the radiolabeled trisaccharide substrate GalNAc4S-GlcA-GalitolNAc4S for 18 h (22). Each fraction isolated from CHOf4S and CHOD520N Percoll gradients was assayed for total arylsulfatase activity using the fluorogenic substrate, 4MUS (18).…”

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Mild Feline Mucopolysaccharidosis Type VI

Yogalingam

Hopwood

Crawley

et al. 1998

Journal of Biological Chemistry

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The missense mutation, L476P, in the N-acetylgalactosamine 4-sulfatase (4S) gene, has previously been shown to be associated with a severe feline mucopolysaccharidosis type VI (MPS VI) phenotype. The present study describes a second mutation, D520N, in the same MPS VI cat colony, which is inherited independently of L476P and is associated with a clinically mild MPS VI phenotype in D520N/L476P compound heterozygous cats. Biochemical and clinical assessment of L476P homozygous, D520N/L476P compound heterozygous, and D520N homozygous cats demonstrated that the entire range of clinical phenotypes, from severe MPS VI, to mild MPS VI, to normal are clustered within a narrow range of residual 4S activity from 0.5% to 4.6% of normal levels. When overexpressed in CHO-KI cells, the secreted form of D520N 4S was inactivated in neutral pH conditions. In addition, intracellular D520N 4S protein was rapidly degraded and corresponded to 37%, 14.5%, and 0.67% of normal 4S protein levels in the microsomal, endosomal, and lysosomal compartments, respectively. However, the specific activity of lysosomal D520N 4S was elevated 22.5-fold when compared with wild-type 4S. These results suggest that the D520N mutation causes a rapid degradation of 4S protein. The effect of this is partially ameliorated as a result of a significant elevation in the specific activity of mutant D520N 4S reaching the lysosomal compartment. The mucopolysaccharidoses (MPS)1 are a group of lysosomal storage disorders that involve the deficiency of specific enzymes required for the degradation of glycosaminoglycans (GAG). MPS VI is characterized by a deficiency of N-acetylgalactosamine 4-sulfatase (4S), which leads to the lysosomal accumulation and urinary excretion of the GAG dermatan sulfate (DS) (1). The severe MPS VI phenotype is characterized by growth retardation, coarse facial features, joint stiffness, corneal clouding, skeletal deformities, and organ and soft tissue involvement. As a result of DS storage in the heart valve and lung, the normal function of these organs is often compromised, leading to early death in affected individuals. The central nervous system does not appear to be affected, even in individuals with clinically severe MPS VI (2).Patients with MPS VI are diagnosed by elevated levels of DS in their urine and a substantial reduction or lack of 4S activity in cells such as peripheral blood leukocytes and cultured skin fibroblasts. The clinical phenotype of MPS VI patients ranges from severe to relatively mild and has generally been shown to correlate with urinary DS and residual 4S activity levels (2). Generally, patients manifest symptomology associated with MPS VI when their 4S protein and activity levels are 5% or less of normal controls (3).Currently, there is no safe and effective treatment available for MPS patients other than surgical intervention to alleviate symptoms whenever possible. MPS VI is a good candidate for both enzyme and gene replacement therapy protocols for a number of reasons. First, the complication of targeting r...

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“…Kidney homogenates from 8-month-old mice were generated and processed as described (18). Aliquots representing 8 ,ug of protein were incubated with the trisaccharide substrate N-acetylgalactosamine 4-sulfate-(1-4)-glucuronic acid-(1-3)-N-acetyl[3H]galactosaminitol-4-sulfate in the presence of 2-acetamido 2-deoxy-D-gluconolactone for 37 h at 37°C.…”

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Targeted disruption of the arylsulfatase B gene results in mice resembling the phenotype of mucopolysaccharidosis VI.

Evers

Säftig²,

Schmidt³

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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Mucopolysaccharidosis VI (MPS VI) is a lysosomal storage disease with autosomal recessive inheritance caused by a deficiency of the enzyme arylsulfatase B (ASB), which is involved in degradation of dermatan sulfate and chondroitin 4-sulfate. A MATERIALS AND METHODSIsolation of a Genomic Clone and Targeting Vector Construction. A 758-bp partial murine ASB cDNA clone (pCABM750) was isolated from a Agtll brain cDNA library (Clontech) by homology screening with a full-length human ASB cDNA probe (4). The nucleotide sequence of pCABM750 was deposited in the GenBank/EMBL data base (accession no. X92096). pCABM750 exhibits 81.8% sequence identity with the human ASB cDNA (4) on nucleotide and 83.1% on deduced amino acid level (data not shown). In EcoRI digested genomic liver DNA of 129/SvJ mice a 9-kbp and a 1.6-kbp DNA fragment are hybridizing with pCABM750. A subgenomic DNA library was constructed by ligation of 8-to 10-kbp EcoRI DNA fragments from genomic liver DNA of 129/SvJ with X-EMBL4 arms and subsequent packaging according to the instructions of the supplier (Stratagene). A 9-kbp genomic clone (pmAB9E, see Fig. la) was isolated from the library by hybridization with pCABM750. For construction of a targeting vector the 6-kbp BamHI DNA fragment from pmAB9E was subcloned into pBlueskript SK-. A linker fragment from the BamHI site to the Sall site of the plasmid vector pUC18 was ligated to the XhoI site at the 5' end of the neomycin (neo) expression cassette of pMClneo poly(A) (ref.13; Stratagene); the neo cassette in pMClneo poly(A) contains a BamHI site at its 3' end. The neo cassette was inserted into the BglII site in exon 5 of the 6-kbp BamHI subclone described above as a BamHI-BamHI restriction fragment resulting in the targeting vector pAB6Bneo (see Fig. la). The insertion of the neo cassette introduces a premature translational stop codon

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Diagnosis of Maroteaux—Lamy syndrome by the use of radiolabelled oligosaccharides as substrates for the determination of arylsulphatase B activity

Cited by 53 publications

References 26 publications

Human liver N-acetylglucosamine-6-sulphate sulphatase. Purification and characterization

Human liver N-acetylglucosamine-6-sulphate sulphatase. Purification and characterization

Mild Feline Mucopolysaccharidosis Type VI

Targeted disruption of the arylsulfatase B gene results in mice resembling the phenotype of mucopolysaccharidosis VI.

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