Kinders R J, Milenkovic A G, Nordin P, Johnson T C
Biochem J. 1980 Sep 15;190(3):605-14. doi: 10.1042/bj1900605.
Glycopeptides were isolated from the cell surfaces of mouse brain cortical tissue that inhibited both cell division and protein synthesis by cells in culture. The protein-synthesis inhibition appeared to affect most cells exposed and was equally effective against glycoprotein and protein synthesis. The inhibition of protein metabolism was independent of mRNA synthesis and uptake of labelled precursors into intracellular pools, indicating that it was directed at intracellular translational events. Fractionation of chloroform/methanol-extracted preparations of this brain cell-surface substance on Bio-Gel P-100 revealed the material to be quite heterogenous, although inhibitory activity was found only in fractions of mol.wt. 25000--30000 and 6000--10000. Biochemical analysis of these fractions demonstrated that they were 6% carbohydrate and 94% amino acid by weight. The 25000--30000-mol.wt. glycopeptides were shown to inhibit cell growth at concentrations of 2 microgram/ml in cultured cells and to inhibit protein synthesis by 50% at concentrations of 3 microgram/ml. The 25000--30000-mol.wt. brain-cell-surface-substance glycopeptides were further purified by ultrafiltration and affinity chromatography with Ulex europaeus agglutinin, resulting in a 400-fold increase in specific biological activity. The inhibitor was not lethal to cells and was not species- or tissue-specific.
从小鼠脑皮质组织的细胞表面分离出了糖肽,这些糖肽可抑制培养细胞的细胞分裂和蛋白质合成。蛋白质合成抑制似乎影响大多数暴露的细胞,并且对糖蛋白和蛋白质合成同样有效。蛋白质代谢的抑制与mRNA合成以及标记前体进入细胞内池无关,这表明它针对的是细胞内的翻译事件。对这种脑细胞表面物质的氯仿/甲醇提取物在Bio-Gel P-100上进行分级分离,结果显示该物质相当不均一,尽管仅在分子量为25000 - 30000和6000 - 10000的级分中发现了抑制活性。对这些级分的生化分析表明,按重量计它们含有6%的碳水化合物和94%的氨基酸。分子量为25000 - 30000的糖肽在培养细胞中浓度为2微克/毫升时可抑制细胞生长,在浓度为3微克/毫升时可抑制50%的蛋白质合成。分子量为25000 - 30000的脑细胞表面物质糖肽通过超滤和用欧洲荆豆凝集素进行亲和层析进一步纯化,其比生物活性提高了400倍。该抑制剂对细胞无致死性,也无物种或组织特异性。