ArticleThe Biochemical journal1997
Expression, purification and characterization of recombinant caprine N-acetylglucosamine-6-sulphatase.
Article in The Biochemical journal, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 34 citations in OpenAlex.
- Tandem mass spectrometric assay of N-acetylglucosamine-6-sulfatase for multiplex analysis of mucopolysaccharidosis-IIID in dried blood spots.Molecular genetics and metabolism · 2024Article
- Evaluation of artificial signal peptides for secretion of two lysosomal enzymes in CHO cells.The Biochemical journal · 2021Article
- Enzyme Replacement Therapy for Mucopolysaccharidosis IIID using Recombinant Human α-Molecular pharmaceutics · 2021Article
- Structural and Mechanistic Analysis of the Choline Sulfatase from Sinorhizobium melliloti: A Class I Sulfatase Specific for an Alkyl Sulfate Ester.Journal of molecular biology · 2018Article
- Neutralizing the anticoagulant activity of ultra-low-molecular-weight heparins using N-acetylglucosamine 6-sulfatase.The FEBS journal · 2013Article
- Sulfatase activities towards the regulation of cell metabolism and signaling in mammals.Cellular and molecular life sciences : CMLS · 2010Review
- Overexpression of inactive arylsulphatase mutants and in vitro activation by light-dependent oxidation with vanadate.The Biochemical journal · 2004Article
- Cloning and characterization of two extracellular heparin-degrading endosulfatases in mice and humans.The Journal of biological chemistry · 2002Article
- Caprine mucopolysaccharidosis IIID: a preliminary trial of enzyme replacement therapy.Journal of molecular neuroscience : MN · 2000Article
- Sequence analysis of heparan sulphate and heparin oligosaccharides.The Biochemical journal · 1999Article
- A strategy for rapid sequencing of heparan sulfate and heparin saccharides.Proceedings of the National Academy of Sciences of the United States of America · 1999Article
- Recombinant caprine 3H-[N-acetylglucosamine-6-sulfatase] and human 3H-[N-acetylgalactosamine-4-sulfatase]: plasma clearance, tissue distribution, and cellular uptake in the rat.Journal of molecular neuroscience : MN · 1998Article
- Recombinant human sulphamidase: expression, amplification, purification and characterization.The Biochemical journal · 1998Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
Mucopolysaccharidosis type IIID or Sanfilippo D syndrome is a lysosomal storage disorder caused by the deficiency of N-acetylglucosamine-6-sulphatase (Glc6S). In addition to human patients, a Nubian goat with this disorder has been described and the caprine Glc6S (cGlc6S) cDNA cloned. In this study, the full-length cGlc6S cDNA was inserted into the expression vector, pEFNeo, which placed the cGlc6S cDNA under the transcriptional control of the human polypeptide chain elongation factor promoter. The pEFNeo expression vector also contains the human growth hormone polyadenylation signal and the genes encoding resistance to ampicillin and G418. The cGlc6S expression construct was electroporated into Chinese hamster ovary (CHO-K1) cells, and stably transfected clones were isolated. One clone, CHOrcGlc6S.17, which secreted the highest Glc6S activity into the culture medium, was selected and cultured in cell factories. The secreted recombinant cGlc6S (rcGlc6S) precursor was purified to homogeneity from conditioned medium by a two-column procedure which consisted of a Cu2+-chelating Sepharose column followed by TSK G3000SW gel filtration. The native molecular mass of rcFlc6S was estimated to be 102 kDa and the subunit size was 94 kDa. The kinetic properties of cGlc6S were similar to those of human Glc6S isolated from liver. rcGlc6S was endocytosed by fibroblasts from patients with mucopolysaccharidosis type IIID via the mannose 6-phosphate receptor-mediated pathway resulting in correction of the storage phenotype of these cells.
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