人核心蛋白聚糖结构域I作为具有20 kDa糖胺聚糖链的重组硫酸乙酰肝素蛋白聚糖的表达。

Expression of human perlecan domain I as a recombinant heparan sulfate proteoglycan with 20-kDa glycosaminoglycan chains.

作者信息

Graham L D, Whitelock J M, Underwood P A

机构信息

CSIRO Molecular Science, Sydney Laboratory, North Ryde, New South Wales, 1670, Australia.

出版信息

Biochem Biophys Res Commun. 1999 Mar 24;256(3):542-8. doi: 10.1006/bbrc.1999.0377.

DOI:10.1006/bbrc.1999.0377

PMID:10080934

Abstract

Recombinant forms of human perlecan domain I were secreted as proteoglycans by stably transfected human 293 cells. A recombinant domain I-only proteoglycan spanned the 95- to 265-kDa region in SDS-PAGE and appeared to be 160 kDa in denaturing gel filtration. Its glycosaminoglycan (GAG) content was approximately 67% heparan sulfate, and its average GAG chain size of 20 kDa suggested that the true molecular mass of the proteoglycan was 90 kDa. Domain I with enhanced green fluorescent protein fused to its C-terminus had an apparent molecular mass of 210-220 kDa and contained approximately 100% heparan sulfate. Its average GAG chain size (also 20 kDa) suggested a true molecular mass of 117 kDa for this proteoglycan. Its sulfate content of 53-77 mol SO2-4 per mole of protein indicated the presence of one sulfate group per 4-7 GAG sugar residues.

摘要

人核心蛋白聚糖结构域I的重组形式由稳定转染的人293细胞作为蛋白聚糖分泌。一种仅含结构域I的重组蛋白聚糖在SDS-PAGE中跨越95至265 kDa区域，在变性凝胶过滤中似乎为160 kDa。其糖胺聚糖（GAG）含量约为67%硫酸乙酰肝素，其平均GAG链大小为20 kDa，表明该蛋白聚糖的真实分子量为90 kDa。在其C末端融合了增强型绿色荧光蛋白的结构域I的表观分子量为210 - 220 kDa，且含有约100%硫酸乙酰肝素。其平均GAG链大小（同样为20 kDa）表明该蛋白聚糖的真实分子量为117 kDa。其每摩尔蛋白质的硫酸盐含量为53 - 77摩尔SO2-4，表明每4 - 7个GAG糖残基存在一个硫酸基团。