Kharchenko E P, Shestak K I, Nalivaeza N N, Sokolova T V
Biokhimiia. 1980 Aug;45(8):1385-98.
Electrophoresis of endonuclease-split rat liver chromatin in polyacrylamide and agarose gels of a monotonous concentration revealed different separation patterns. The first pattern exhibited a spectrum of chromatin fragments of discrete sizes, while the second one -- two fractions, i. e. DNP and DNA. The polyacrylamide gel concentration gradient allowed to achieve the best resolution of individual classes of chromation fragments with individual subfractions in sub-, mono- and trinucleosomes. A comparison of micrococcal nuclease-split chromatin fragments of vertebrates revealed nucleosomes of varying sizes. The heterogeneity of the nucleosomes was also found during isotachophoresis and free flow electrophoresis. An analysis of individual subfractions of the mononucleosomes revealed differences in the composition and number of proteins as well as in the kinetics of basic and acidic titration. The chromatin fractions obtained by discontinuous endonucleolysis differed in their composition from the individual subfractions of histones.
在浓度单一的聚丙烯酰胺和琼脂糖凝胶中对经核酸内切酶切割的大鼠肝脏染色质进行电泳,结果显示出不同的分离模式。第一种模式呈现出一系列大小离散的染色质片段,而第二种模式则是两个组分,即脱氧核蛋白(DNP)和DNA。聚丙烯酰胺凝胶浓度梯度能够实现对单个类别的染色质片段以及亚核小体、单核小体和三核小体中的各个亚组分的最佳分辨率。对脊椎动物微球菌核酸酶切割的染色质片段进行比较,发现了大小各异的核小体。在等速电泳和自由流动电泳过程中也发现了核小体的异质性。对单核小体的各个亚组分进行分析,揭示了蛋白质组成和数量以及酸碱滴定动力学方面的差异。通过不连续核酸内切酶解获得的染色质组分在组成上与组蛋白的各个亚组分不同。