Mishima K, Tsuchiya M, Tanigawa Y, Yoshimura Y, Shimoyama M
Department of Biochemistry, Shimane Medical University, Japan.
Eur J Biochem. 1989 Feb 1;179(2):267-73. doi: 10.1111/j.1432-1033.1989.tb14550.x.
A non-histone acceptor protein for hen liver nuclear ADP-ribosyltransferase was purified to an apparently homogeneous state through salt extraction and chromatography on hydroxyapatite, phenyl-Sepharose, carboxy-methyl-cellulose, Sephadex G-75, phenyl 5-PW, mono S and Radial PAK C18. This protein was termed p33. The ADP-ribosylation of p33 was enhanced more than 60-fold by double-stranded DNA. Single-stranded DNA, RNA and poly(L-glutamate), but not deoxyribonucleotide, were partially effective. DNA-dependent ADP-ribosylation was also observed when whole histones were used as acceptor. DNA required for the maximal ADP-ribosylation depended on the dose of the acceptor protein; the optimal mass ratio of DNA to the acceptor protein was 1:1 with both p33 and whole histones. DNA decreased the Km for NAD and concomitantly increased the Vmax value, but did not alter the Km for p33. These results are consistent with the idea that p33 may participate in chromatin processes such as replication or transcription, through modification by nuclear ADP-ribosyltransferase.
通过盐提取以及在羟基磷灰石、苯基 - 琼脂糖、羧甲基纤维素、葡聚糖G - 75、苯基5 - PW、单S和径向PAK C18上进行层析，将鸡肝细胞核ADP - 核糖基转移酶的一种非组蛋白受体蛋白纯化至明显的均一状态。这种蛋白被命名为p33。双链DNA使p33的ADP - 核糖基化增强了60倍以上。单链DNA、RNA和聚（L - 谷氨酸）有部分作用，但脱氧核苷酸则无作用。当使用全组蛋白作为受体时，也观察到了DNA依赖性的ADP - 核糖基化。最大ADP - 核糖基化所需的DNA量取决于受体蛋白的剂量；DNA与受体蛋白的最佳质量比对于p33和全组蛋白均为1:1。DNA降低了NAD的Km值，同时增加了Vmax值，但未改变p33的Km值。这些结果与p33可能通过核ADP - 核糖基转移酶的修饰参与染色质过程（如复制或转录）的观点一致。
