Expression, purification and characterization of recombinant human N-acetylgalactosamine-6-sulphatase

J. Bielicki, M. Fuller, X. H. Guo, C. P. Morris, J. J. Hopwood, D. S. Anson

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33 Citations (Scopus)


Full-length cDNA sequences encoding human N-acetylgalactosamine-6-sulphatase were stably expressed in Chinese hamster ovary cells under the transcriptional control of the human polypeptide chain elongation factor 1α gene promoter. A clonal cell line overexpressing recombinant N-acetylgalactosamine-6-sulphatase to a level of approx. 3 mg/1 of culture medium was isolated. The secreted precursor enzyme was purifed to homogeneity by a two-column procedure with an overall yield of 53% of the activity. The physical and catalytic parameters of the recombinant enzyme were similar to those of the mature form isolated from liver. On SDS/PAGE and gel filtration, recombinant N-acetylgalactosamine-6-sulphatase had a native molecular mass of 58-60 kDa. Recombinant N-acetylgalactosamine-amine-6-sulphatase was endocytosed by mucopolysaccharidosis IVA fibroblasts via the mannose-6-phosphate receptor-mediated pathway and was efficiently localized to lysosomes.

Original languageEnglish
Pages (from-to)333-339
Number of pages7
JournalBiochemical Journal
Issue number1
Publication statusPublished or Issued - 1995

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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