Schmidtchen A, Sundler R, Fransson L A
Department of Physiological Chemistry, University of Lund, Sweden.
Glycoconj J. 1990;7(6):563-72. doi: 10.1007/BF01189077.
Here we present evidence that a fibroblast heparan sulphate proteoglycan of approx. 300 kDa and with a core protein of apparent molecular mass 70 kDa is covalently linked to the plasma membrane via a linkage structure involving phosphatidylinositol. Phosphatidylinositol-specific phospholipase C releases such a heparan sulphate proteoglycan only from cells labelled with [35S]sulphate in the absence of serum. Cell cultures labelled with [3H]myo-inositol in the absence or presence of serum produce a radiolabelled heparan sulphate proteoglycan which was purified by gel-permeation chromatography and ion-exchange chromatography on MonoQ. Digestion with heparan sulphate lyase and analysis by gel-permeation chromatography and sodium dodecylsulphate-polyacrylamide gel-electrophoresis revealed that the 3H-label is associated with a core protein of apparent mass 70 kDa.
在此,我们提供证据表明,一种约300 kDa的成纤维细胞硫酸乙酰肝素蛋白聚糖,其核心蛋白表观分子量为70 kDa,通过涉及磷脂酰肌醇的连接结构与质膜共价连接。磷脂酰肌醇特异性磷脂酶C仅在无血清条件下从用[35S]硫酸盐标记的细胞中释放出这种硫酸乙酰肝素蛋白聚糖。在无血清或有血清条件下用[3H]肌醇标记的细胞培养物产生一种放射性标记的硫酸乙酰肝素蛋白聚糖,该蛋白聚糖通过凝胶渗透色谱和MonoQ离子交换色谱进行纯化。用硫酸乙酰肝素裂解酶消化并通过凝胶渗透色谱和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明,3H标记与表观质量为70 kDa的核心蛋白相关。