Surowy C S, Berger N A
J Biol Chem. 1983 Jan 10;258(1):579-83.
The effect of diadenosine 5', 5"'-P1,P4-tetraphosphate (Ap4A) on the time course and acceptors of poly(ADP-ribose) synthesis was studied in undamaged and N-methyl-N'-nitro-N-nitrosoguanidine-treated human lymphocytes. Analysis of protein acceptors of poly(ADP-ribose) revealed that treatment with Ap4A stimulated ADP-ribosylation of bands at molecular weights of 96,000, 79,000, and 62,000. Pulse-chase studies showed that these bands were produced as a result of an effect of Ap4A on the processing of ADP-ribosylated proteins rather than on the synthesis of newly ADP-ribosylated proteins. By incubating permeabilized cells in the absence or presence of Ap4A and purified poly(ADP-ribose) polymerase auto-ADP-ribosylated with [32P]NAD+, we showed that the Mr = 96,000, 79,000, and 62,000 bands were derivatives of the prelabeled enzyme. Our results indicate that normal human lymphocytes process auto-ADP-ribosylated poly(ADP-ribose) polymerase to specific lower molecular weight products and that this processing is stimulated by Ap4A.
在未受损及经N-甲基-N'-硝基-N-亚硝基胍处理的人淋巴细胞中,研究了5',5'''-P1,P4-四磷酸二腺苷(Ap4A)对聚(ADP-核糖)合成的时间进程及受体的影响。对聚(ADP-核糖)的蛋白质受体分析显示,用Ap4A处理可刺激分子量为96,000、79,000和62,000条带的ADP-核糖基化。脉冲追踪研究表明,这些条带是Ap4A对ADP-核糖基化蛋白质加工的影响所致,而非对新的ADP-核糖基化蛋白质合成的影响。通过在有无Ap4A及用[32P]NAD+进行自身ADP-核糖基化的纯化聚(ADP-核糖)聚合酶存在的情况下孵育通透细胞,我们发现分子量为96,000、79,000和62,000的条带是预标记酶的衍生物。我们的结果表明,正常人淋巴细胞将自身ADP-核糖基化的聚(ADP-核糖)聚合酶加工成特定的低分子量产物,且这种加工受到Ap4A的刺激。