EXPRESSION PURIFICATION AND CHARACTERIZATION OF RECOMBINANT HUMAN N-ACETYLGALACTOSAMINE-6-SULFATASE

被引：32

作者：

BIELICKI, J ^{[1
]}

FULLER, M ^{[1
]}

GUO, XH ^{[1
]}

MORRIS, GP ^{[1
]}

HOPWOOD, JJ ^{[1
]}

ANSON, DS ^{[1
]}

机构：

[1] WOMENS & CHILDRENS HOSP,DEPT CHEM PATHOL,LYSOSOMAL DIS RES UNIT,ADELAIDE,SA 5006,AUSTRALIA

来源：

BIOCHEMICAL JOURNAL | 1995年 / 311卷

关键词：

D O I：

10.1042/bj3110333

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Full-length cDNA sequences encoding human N-acetyl-galactosamine-6-sulphatase were stably expressed in Chinese hamster ovary cell under the transcriptional control of the human polypeptide chain elogation 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-sulphtase was endocytosed by mucopolysaccharidosis IVA fibroblasts via the mannose-6-phosphate receptor-mediated pathway and was efficiently localized to lysosomes.

引用

页码：333 / 339

页数：7

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