Garg H G, Lippay E W, Neame P J
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA.
Carbohydr Res. 1992 Jan;223:209-20. doi: 10.1016/0008-6215(92)80017-u.
Proteoglycans (PGs) from human burn hypertrophic scar of a patient with Ehlers-Danlos syndrome were extracted with 4M guanidinium chloride and purified by DEAE-cellulose chromatography. Differential ethanol precipitation of the PG fraction obtained after ion-exchange chromatography yielded two low mol.-wt. PGs, on rich in glucuronic acid (PGGLCA; Mr 66 kDa) and the other rich in iduronic acid (PGIDOA; Mr 48 kDa). In PGGLCA, 84% of the glycosaminoglycan chains are composed of GlcA----GalNAc(SO4) units, whereas in PGIDOA, the chains contain 95% IdoA----GalNAc(SO4) disaccharide units. Upon treatment with testicular hyaluronidase, the PGs gave different-sized oligosaccharides. Chondroitinase ABC digestion of PGGLCA or PGIDOA gave a single protein core (Mr approximately 20 kDa). The presence of glucosamine and sialic acid in PGGLCA and PGIDOA suggests that both contain N-linked oligosaccharides.
采用4M氯化胍从一名患有埃勒斯-当洛综合征患者的人烧伤增生性瘢痕中提取蛋白聚糖(PGs),并通过DEAE-纤维素色谱法进行纯化。离子交换色谱后获得的PG级分经不同浓度乙醇沉淀,得到两种低分子量的PGs,一种富含葡萄糖醛酸(PGGLCA;分子量66 kDa),另一种富含艾杜糖醛酸(PGIDOA;分子量48 kDa)。在PGGLCA中,84%的糖胺聚糖链由GlcA----GalNAc(SO4)单元组成,而在PGIDOA中,链中含有95%的IdoA----GalNAc(SO4)二糖单元。经睾丸透明质酸酶处理后,PGs产生了不同大小的寡糖。用软骨素酶ABC消化PGGLCA或PGIDOA得到单一的蛋白核心(分子量约20 kDa)。PGGLCA和PGIDOA中存在葡萄糖胺和唾液酸表明两者都含有N-连接寡糖。