Hamilton T A, Sussman H H
Biochem J. 1981 Jul 15;198(1):29-35. doi: 10.1042/bj1980029.
The effect of 5-bromo-2'-deoxyuridine (BrdUrd) and dibutyryl cyclic AMP (Bt2cAMP) on the expression of the placental isoenzyme of human alkaline phosphatase was examined in BeWo choriocarcinoma cells. By using a combination of specific immunoprecipitation and polyacrylamide-gel electrophoresis of cells labelled either metabolically with [35S]methionine or cell-surface-labelled with 125I, both BrdUrd (5 micrograms/ml) and 1 mM-Bt2cAMP were shown to result in the enhanced accumulation of a specific protein. This protein has immunochemical identity and co-electrophoreses with placental alkaline phosphatase in two-dimensional gels. These results clearly demonstrate that the induction of placental alkaline phosphatase activity in choriocarcinoma cells treated with these agents is a consequence of the accumulation of specific enzyme protein rather than of altered catalytic activity.
在BeWo绒毛膜癌细胞中检测了5-溴-2'-脱氧尿苷(BrdUrd)和二丁酰环磷腺苷(Bt2cAMP)对人碱性磷酸酶胎盘同工酶表达的影响。通过结合使用用[35S]甲硫氨酸进行代谢标记或用125I进行细胞表面标记的细胞的特异性免疫沉淀和聚丙烯酰胺凝胶电泳,结果显示,BrdUrd(5微克/毫升)和1毫摩尔/升的Bt2cAMP均导致一种特定蛋白质的积累增加。这种蛋白质具有免疫化学同一性,并且在二维凝胶中与胎盘碱性磷酸酶共电泳。这些结果清楚地表明,在用这些试剂处理的绒毛膜癌细胞中,胎盘碱性磷酸酶活性的诱导是特定酶蛋白积累的结果,而不是催化活性改变的结果。