Itzhaki R F
Biochem J. 1971 May;122(4):583-92. doi: 10.1042/bj1220583.
The binding of deoxyribonucleoprotein to Toluidine Blue, to cetylpyridinium chloride and to polylysine of various molecular weights was studied to determine the percentage of free DNA phosphate groups in deoxyribonucleoprotein. Binding was measured by addition of these reagents to deoxyribonucleoprotein at a range of concentrations such that complete precipitation of the deoxyribonucleoprotein occurred. With Toluidine Blue the binding corresponded to about 48% of the DNA phosphates in deoxyribonucleoprotein. The dye did not cause appreciable displacement of protein from the DNA. With cetylpyridinium chloride the binding corresponded to about 41% of the DNA phosphates. With polylysine preparations of molecular weight 1250 and 7790 the binding values for deoxyribonucleoprotein were 46 and 38% respectively. The results suggest that the free phosphates lie in stretches sufficiently long to accommodate most of each polylysine molecule. With polylysine of molecular weight 62000 cross-linking of free stretches of DNA on different deoxyribonucleoprotein molecules probably occurs. It is concluded that although most of the free phosphates are probably ;hidden' beneath covering histone, corresponding perhaps to runs of non-basic residues in the latter, they are surprisingly accessible to very large molecules. The relevance of this finding to the problem of gene repression is discussed.
研究了脱氧核糖核蛋白与甲苯胺蓝、十六烷基氯化吡啶以及不同分子量的聚赖氨酸的结合情况,以确定脱氧核糖核蛋白中游离DNA磷酸基团的百分比。通过在一系列浓度下将这些试剂添加到脱氧核糖核蛋白中来测量结合情况,使得脱氧核糖核蛋白完全沉淀。对于甲苯胺蓝,结合量相当于脱氧核糖核蛋白中约48%的DNA磷酸盐。该染料不会导致蛋白质从DNA上明显位移。对于十六烷基氯化吡啶,结合量相当于约41%的DNA磷酸盐。对于分子量为1250和7790的聚赖氨酸制剂,脱氧核糖核蛋白的结合值分别为46%和38%。结果表明,游离磷酸盐所在的片段足够长,能够容纳每个聚赖氨酸分子的大部分。对于分子量为62000的聚赖氨酸,不同脱氧核糖核蛋白分子上的游离DNA片段可能发生交联。得出的结论是,尽管大多数游离磷酸盐可能“隐藏”在覆盖的组蛋白之下,这可能对应于组蛋白中连续的非碱性残基,但它们对于非常大的分子来说却出人意料地易于接近。讨论了这一发现与基因抑制问题的相关性。