Caputo C B, Kimura J H, Hascall V C
Arch Biochem Biophys. 1984 May 1;230(2):594-604. doi: 10.1016/0003-9861(84)90440-5.
Rat chondrosarcoma chondrocytes were cultured in the presence of puromycin to induce premature termination of core protein precursor. The structure and function of intracellular and extracellular proteoglycans were assessed by molecular sieve chromatography and polyacrylamide gel electrophoresis. [3H]Serine incorporation was maximally inhibited by 3 X 10(-4) M puromycin but unaffected by 10(-5) M puromycin. Proteoglycans synthesized in the presence of puromycin exhibited increased monomer size due to increased chondroitin sulfate chain size, typical of proteoglycans synthesized in the presence of protein synthesis inhibitors, but no loss in ability to bind to hyaluronic acid; and no loss in core protein size was observed after treatment with chondroitinase. These data suggest that chondrocytes select only completed or nearly completed core protein molecules to process into proteoglycans.
在嘌呤霉素存在的情况下培养大鼠软骨肉瘤软骨细胞,以诱导核心蛋白前体的提前终止。通过分子筛色谱法和聚丙烯酰胺凝胶电泳评估细胞内和细胞外蛋白聚糖的结构和功能。[3H]丝氨酸掺入在3×10(-4)M嘌呤霉素作用下受到最大抑制,但不受10(-5)M嘌呤霉素影响。在嘌呤霉素存在下合成的蛋白聚糖由于硫酸软骨素链大小增加而表现出单体大小增加,这是在蛋白质合成抑制剂存在下合成的蛋白聚糖的典型特征,但与透明质酸结合的能力没有丧失;用软骨素酶处理后未观察到核心蛋白大小的损失。这些数据表明,软骨细胞仅选择完整或接近完整的核心蛋白分子来加工成蛋白聚糖。