人胚肺成纤维细胞株纤连蛋白结构域N─糖链结构研究

本文应用明胶、肝素亲和层析二步法首先纯化了人胚肺成纤维细胞培养液的纤连蛋白（Ｆｉｂｒｏｎｅｏｔｈｅ，Ｆｎ），经ＳＤＳ－ＰＡＧＥ鉴定为一条带，然后用胰糜蛋白酶消化纯化的Ｆｎ所获得的酶解波，经分离分别得到明胶结合片段和肝素结合片段，再应用凝集素－ＨＲＰ染色的Ｗｅｓｔｅｎ转移电泳法研究糖链结构，结果证实：１．Ｆｎ中明胶结合片段（４４ｋｄ）中含有二天线和多天线复杂型糖链，并接有平分型ｇｌｃＮＡｃ糖基、核心力Ｆｕｃ。２肝素结合片段（３０ｋｄ）只含有二天线复杂型糖链，不含平分型ＧｌｃＮＡｃ糖基及核心Ｆｕｃ．

Study on the Structure of N-Linked Oligosaccharide of Purified Fibronectin Domains from Human Embryo Lung Fibroblast

Fibroneotin(Fn)was purified from cultured human embryo lung fibroblast by affinity chromatography.It appeared as a single band on SDS-PAGE.Both gelatin-binding and heparin-binding fragments were obtained separately from the chymotrypsinolysate of purified Fn by special affinity columns.Based on the speciflcity,in recognition of lectins for the oligosaocharides,western blot electrophoresis with the lectin-HRP staining was applied to analyse the strueture of the oligosacchrides in the gelatin-binding and the heparin-binding fragments.The results suggested that:1.Gelatin-binding fragment(44kd)of human embryo lung fibroblast Fn eontains not only complex type oligosaccharides(including bi-,tri- and tetraantenery ones)but also biseoling GlcNAc and core fucose.2.Heparin-binding fragment(30kd)has only biantenery complex type oligossaoharide without bisecting GlcNAc and core fucose.