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. 1995 Oct 1;311(Pt 1):333–339. doi: 10.1042/bj3110333

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

J Bielicki 1, M Fuller 1, X H Guo 1, C P Morris 1, J J Hopewood 1, D S Anson 1
PMCID: PMC1136156  PMID: 7575473

Abstract

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 1 alpha gene promoter. A clonal cell line overexpressing recombinant N-acetylgalactosamine-6-sulphatase to a level of approx. 3 mg/l of culture medium was isolated. The secreted precursor enzyme was purified 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-6-sulphatase was endocytosed by mucopolysaccharidosis IVA fibroblasts via the mannose-6-phosphate receptor-mediated pathway and was efficiently localized to lysosomes.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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