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https://hdl.handle.net/2440/7020
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dc.contributor.author | Bielicki, J. | - |
dc.contributor.author | Fuller, M. | - |
dc.contributor.author | Guo, X.H. | - |
dc.contributor.author | Morris, C. | - |
dc.contributor.author | Hopwood, J. | - |
dc.contributor.author | Anson, D. | - |
dc.date.issued | 1995 | - |
dc.identifier.citation | Biochemical Journal, 1995; 311(1):333-339 | - |
dc.identifier.issn | 0264-6021 | - |
dc.identifier.issn | 1470-8728 | - |
dc.identifier.uri | http://hdl.handle.net/2440/7020 | - |
dc.description.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. | - |
dc.language.iso | en | - |
dc.publisher | Biochemical Society | - |
dc.source.uri | http://dx.doi.org/10.1042/bj3110333 | - |
dc.subject | Liver | - |
dc.subject | CHO Cells | - |
dc.subject | Fibroblasts | - |
dc.subject | Animals | - |
dc.subject | Humans | - |
dc.subject | Mucopolysaccharidosis IV | - |
dc.subject | Chondroitinsulfatases | - |
dc.subject | Recombinant Proteins | - |
dc.subject | DNA, Complementary | - |
dc.subject | Culture Media, Conditioned | - |
dc.subject | Gene Transfer Techniques | - |
dc.subject | Endocytosis | - |
dc.subject | Gene Expression | - |
dc.subject | Base Sequence | - |
dc.subject | Kinetics | - |
dc.subject | Genetic Vectors | - |
dc.subject | Molecular Sequence Data | - |
dc.subject | Cricetinae | - |
dc.title | Expression, purification and characterization of recombinant human N-acetylgalactosamine-6-sulphatase | - |
dc.type | Journal article | - |
dc.identifier.doi | 10.1042/bj3110333 | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | Fuller, M. [0000-0001-9092-8942] | - |
Appears in Collections: | Aurora harvest 5 Paediatrics publications |
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