Greyling H J, Sewell B T, Von Holt C
Department of Biochemistry, University of Cape Town, Rondebosch, South Africa.
Eur J Biochem. 1988 Feb 1;171(3):721-6. doi: 10.1111/j.1432-1033.1988.tb13845.x.
A hybrid histone octamer was reconstituted from erythrocyte H2A and H2B, avian [110 Cys-des-thio]histone H3 and the sea-urchin sperm [73Cys]H4 variant. [110Cys-Des-thio]histone H3 was prepared by reaction of natural H3 with Raney nickel. The ability of the hybrid octamer to crystallize to the same form as the natural octamer demonstrated that the chemical modification of cysteine to alanine in H3 and the mutation from threonine to cysteine in sperm H4 do not alter histone-histone interactions in the octamer. Since the sulfhydryl groups of both H4 molecules are fully accessible to 5,5'-dithiobis(2-nitrobenzoate) these residues provide suitable sites for the introduction of a single cysteine-specific label per H4 molecule in the octamer.
一种杂合组蛋白八聚体由红细胞H2A和H2B、禽[110半胱氨酸-去硫代]组蛋白H3和海胆精子[73半胱氨酸]H4变体重构而成。[110半胱氨酸-去硫代]组蛋白H3通过天然H3与雷尼镍反应制备。杂合八聚体结晶成与天然八聚体相同形式的能力表明，H3中半胱氨酸向丙氨酸的化学修饰以及精子H4中苏氨酸向半胱氨酸的突变不会改变八聚体中组蛋白-组蛋白的相互作用。由于两个H4分子的巯基都能完全被5,5'-二硫代双（2-硝基苯甲酸）接触，这些残基为在八聚体中每个H4分子引入单个半胱氨酸特异性标记提供了合适的位点。
