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

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