Inoue A, Higashi Y, Hasuma T, Morisawa S, Yukioka M
Eur J Biochem. 1983 Sep 1;135(1):61-8. doi: 10.1111/j.1432-1033.1983.tb07617.x.
A set of non-histone proteins has been identified in the nuclei from liver, brain, spleen and testis tissues of the rat. Following moderate digestion of thoroughly washed nuclei with DNase I or micrococcal nuclease, EDTA was added to 5 mM to the reaction mixture and the preparation centrifuged. We found that the supernatant contained a limited amount of non-histone proteins (fraction S1). Sodium dodecyl sulfate (SDS) gel electrophoresis revealed S1 to be composed of a remarkably simple set of proteins resolved into four groups (A-D) each possessing closely spaced doublets or a triplet. Their molecular weights were A, 76 100-80 000; B, 48 200-49 500; C, 44 500-45 200 and D, 39 500-41 500. The yield suggested that these proteins were structural constituents; however, they did not coincide with the known structural proteins of the cell nucleus. Two-dimensional gel electrophoresis further resolved each of the SDS bands into as many as nine spots, according to various charges. Some were labelled with [32P]orthophosphate in vivo, or with [gamma-32P]ATP and purified nuclear protein kinase NII in vitro. The released proteins B-D had fairly constant relative molar ratios at various times of digestion, thereby indicating possible localizations at similar sites in the nucleus. The kinetic data together with the aggregation property at neutral pH values and the solubility in 5 mM EDTA suggest that proteins B-D constitute a group of proteins that have several common characteristics.
在大鼠肝脏、大脑、脾脏和睾丸组织的细胞核中已鉴定出一组非组蛋白。用脱氧核糖核酸酶I或微球菌核酸酶对充分洗涤过的细胞核进行适度消化后，向反应混合物中加入5 mM的乙二胺四乙酸（EDTA），然后对制备物进行离心。我们发现，上清液中含有少量非组蛋白（S1组分）。十二烷基硫酸钠（SDS）凝胶电泳显示，S1由一组非常简单的蛋白质组成，可分为四组（A - D），每组都有紧密间隔的双峰或三峰。它们的分子量分别为：A，76100 - 80000；B，48200 - 49500；C，44500 - 45200；D，39500 - 41500。产量表明这些蛋白质是结构成分；然而，它们与已知的细胞核结构蛋白并不一致。二维凝胶电泳根据不同电荷进一步将每个SDS条带解析为多达九个斑点。其中一些在体内用[32P]正磷酸盐标记，或在体外用[γ-32P]ATP和纯化的核蛋白激酶NII标记。释放的蛋白质B - D在不同消化时间具有相当恒定的相对摩尔比，从而表明它们可能定位于细胞核中的相似位点。动力学数据以及在中性pH值下的聚集特性和在5 mM EDTA中的溶解度表明，蛋白质B - D构成了一组具有几个共同特征的蛋白质。
