Moss T, Cary P D, Abercrombie B D, Crane-Robinson C, Bradbury E M
Eur J Biochem. 1976 Dec 11;71(2):337-50. doi: 10.1111/j.1432-1033.1976.tb11120.x.
It has been shown by high-resolution proton magnetic resonance (PMR) spectroscopy and circular dichroism (CD) that an H2A/H2B histone complex exists after salt extraction of these histones from chromatin and that this complex can be fully renatured from both urea-denatured acid-extracted and from urea-denatured salt-extracted histones. The histone complex is shown to involve specific secondary and tertiary structure. Formation of this complex is observed to be critically dependent on pH, occurring at and above pH 5. It cannot be induced below pH 5 by increase in ionic strength. From CD spectra the H2A/H2B complex is shown to contain about 37% alpha helix but no beta structure, the latter being confirmed by infrared spectroscopy in the 6-mum region. The PMR spectra show that the structured region includes most of the aromatic residues of both histones, at least two histidine residues of H2B and probably histidines 31 and 82 of histone H2A. The secondary structure of histones H2A and H2B is predicted using the Chou and Fasman procedure and comparisons are made between the predictions for histones of different species. These results in conjunction with the experimental evidence lead to the conclusion that at least residues 31-95 of H2A and residues 37-114 of H2B, i.e. the more apolar regions of the molecules, are involved in the tertiary structure of the H2A/H2B complex.
高分辨率质子磁共振(PMR)光谱法和圆二色性(CD)表明,从染色质中盐提取这些组蛋白后存在H2A/H2B组蛋白复合体,并且该复合体可以从尿素变性的酸提取组蛋白和尿素变性的盐提取组蛋白中完全复性。已表明该组蛋白复合体涉及特定的二级和三级结构。观察到该复合体的形成严重依赖于pH值,在pH 5及以上时发生。在pH 5以下,通过增加离子强度不能诱导其形成。从CD光谱可知,H2A/H2B复合体含有约37%的α螺旋,但没有β结构,后者在6μm区域通过红外光谱得到证实。PMR光谱表明,结构化区域包括两种组蛋白的大部分芳香族残基、H2B的至少两个组氨酸残基以及可能的组蛋白H2A的组氨酸31和82。使用Chou和Fasman方法预测了组蛋白H2A和H2B的二级结构,并对不同物种组蛋白的预测结果进行了比较。这些结果与实验证据相结合,得出结论:至少H2A的31-95残基和H2B的37-114残基,即分子中极性较小的区域,参与了H2A/H2B复合体的三级结构。