Khrapunov S N, Dragan A I, Protas A F, Berdyshev G D
Mol Biol (Mosk). 1985 Jul-Aug;19(4):1011-20.
Structure of the (H2A-H2B-H3-H4)2 histone octamer isolated from calf thymus chromatin at ionic strength 0.1 to 4.0 M NaCl, pH 7.6, was studied spectrofluorometrically. Sensitivity of lambda max tyrosine fluorescence position to structural changes of histone oligomers and to the processes of their association was shown. It were detect two ranges of cooperative changes in histone optical parameters at 0.6-1.4 M NaCl (transition I) and at 2.4-3.4 M NaCl (transition II): Transition I corresponds to the formation of equilibrium system (hexamer) + (dimer) in equilibrium octamer. Transition II corresponds to the structural changes of the histone octamer. Thus, fluorescence anisotropy increases, lambda max for fluorescence spectrum is shifted to the longer wavelengths, contributions of two components to fluorescence decay change, a fraction of fluorescence accessible to the quenching by I- decreases. Histone octamer formation is characterized by making specific contacts between the (H2A-H2B) dimer and (H3-H4)2 tetramer. These contacts are realized at gradual changing of ionic strengths (by dialysis). In the case of abrupt local changes of the environment the process is irreversibly shifted to formation of unspecific high molecular aggregates. The important function role for energetically degenerated states of histone oligomers, energy barriers between which can be overcome by changing total conditions of histone microenvironment in chromatin is discussed.
在离子强度为0.1至4.0 M NaCl、pH 7.6的条件下,对从小牛胸腺染色质中分离出的(H2A - H2B - H3 - H4)2组蛋白八聚体的结构进行了荧光光谱研究。研究表明,λmax酪氨酸荧光位置对组蛋白寡聚体的结构变化及其缔合过程具有敏感性。在0.6 - 1.4 M NaCl(转变I)和2.4 - 3.4 M NaCl(转变II)时,检测到组蛋白光学参数的两个协同变化范围:转变I对应于平衡八聚体中平衡体系(六聚体)+(二聚体)的形成。转变II对应于组蛋白八聚体的结构变化。因此,荧光 anisotropy增加,荧光光谱的λmax向更长波长移动,两个组分对荧光衰减的贡献发生变化,可被I -猝灭的荧光部分减少。组蛋白八聚体的形成特征在于(H2A - H2B)二聚体和(H3 - H4)2四聚体之间形成特定接触。这些接触是在离子强度逐渐变化(通过透析)时实现的。在环境发生突然局部变化的情况下,该过程不可逆地转变为形成非特异性高分子聚集体。讨论了组蛋白寡聚体能量简并态的重要功能作用,通过改变染色质中组蛋白微环境的总体条件可以克服它们之间的能量障碍。