Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/7020
Citations | ||
Scopus | Web of ScienceĀ® | Altmetric |
---|---|---|
?
|
?
|
Type: | Journal article |
Title: | Expression, purification and characterization of recombinant human N-acetylgalactosamine-6-sulphatase |
Author: | Bielicki, J. Fuller, M. Guo, X.H. Morris, C. Hopwood, J. Anson, D. |
Citation: | Biochemical Journal, 1995; 311(1):333-339 |
Publisher: | Biochemical Society |
Issue Date: | 1995 |
ISSN: | 0264-6021 1470-8728 |
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. |
Keywords: | Liver CHO Cells Fibroblasts Animals Humans Mucopolysaccharidosis IV Chondroitinsulfatases Recombinant Proteins DNA, Complementary Culture Media, Conditioned Gene Transfer Techniques Endocytosis Gene Expression Base Sequence Kinetics Genetic Vectors Molecular Sequence Data Cricetinae |
DOI: | 10.1042/bj3110333 |
Published version: | http://dx.doi.org/10.1042/bj3110333 |
Appears in Collections: | Aurora harvest 5 Paediatrics publications |
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.