Tsai Y H, Ansevin A T, Hnilica L S
Biochemistry. 1975 Mar 25;14(6):1257-65. doi: 10.1021/bi00677a026.
First derivative thermal denaturation profiles were compared for chromatin samples prepared from chicken erythrocytes, chicken liver, and sea urchin (Strongylocentrotus purpuratus) sperm. Selective dissociation of various histone fractions, including tissue-specific F2c and gamma histones, was manifested in characteristic changes of the thermal denaturation profiles. It was concluded that the binding of individual histone fractions to the DNA can be identified with paricular temperatures of thermal denaturation. This observation was tested by denaturation experiments on hybrid chromatins. Chicken erythrocyte chromatin devoid of F2c histone and reconstituted with isolated liver F1 histone denatured like chicken liver chromatin. Conversely, chicken liver chromatin devoid of F1 histones and reconstituted with isolated F2c fraction exhibited a thermal denaturation profile characteristic of the chicken erythrocyte chromatin. In other words, the thermal denaturation profile of reconstituted chromatin was determined approximately by the sum of contribution histone fractions. The obvious distinctions recognized among the derivative thermal denaturation profiles of compositionally different nucleoproteins suggest that thermal denaturation sensitively detects variations in histone content and therefore is a valuable tool for the routine characterization of chromatin preparations.
比较了从鸡红细胞、鸡肝和海胆(紫球海胆)精子制备的染色质样品的一阶热变性曲线。包括组织特异性F2c和γ组蛋白在内的各种组蛋白组分的选择性解离表现为热变性曲线的特征性变化。得出的结论是,单个组蛋白组分与DNA的结合可以通过特定的热变性温度来识别。通过对杂交染色质的变性实验验证了这一观察结果。不含F2c组蛋白并用分离的肝F1组蛋白重构的鸡红细胞染色质,其变性情况与鸡肝染色质相似。相反,不含F1组蛋白并用分离的F2c组分重构的鸡肝染色质呈现出鸡红细胞染色质特有的热变性曲线。换句话说,重构染色质的热变性曲线大约由组蛋白组分的贡献总和决定。在组成不同的核蛋白的衍生热变性曲线中识别出的明显差异表明,热变性能够灵敏地检测组蛋白含量的变化,因此是染色体制剂常规表征的一种有价值的工具。