Klevan L, Wang J C
Biochemistry. 1980 Nov 11;19(23):5229-34. doi: 10.1021/bi00564a012.
Staphylococcal nuclease digestion of the complex between DNA and DNA gyrase yields a gyrase-DNA core particle composed of a 140 base pair DNA segment and an active gyrase enzyme. The partial specific volume and S20,w of this purified core complex are measured to be 0.70 cm3/g and 14.5 S, respectively, by sedimentation measurements in H2O and D2O media. The molecular weight of the core complex estimated from equilibrium centrifugation is 470 000; the ratio of the translational frictional coefficient to that of the unsolvated equivalent sphere is calculated to be 1.9. Treatment of free gyrase in solution with dimethyl suberimidate gives three cross-linked species of roughly equal amounts that can be identified as alpha 2, alpha 2 beta, and alpha 2 beta 2. When the gyrase core complex is treated with the same cross-linking agent, 70-80% of the protein is converted to the alpha 2 beta 2 species. These results establish that the gyrase-DNA core complex contains a 140 base pair DNA segment and a tetrameric alpha 2 beta 2 protein.
用葡萄球菌核酸酶消化DNA与DNA促旋酶之间的复合物,可产生一种促旋酶-DNA核心颗粒,它由一段140个碱基对的DNA片段和一种有活性的促旋酶组成。通过在H₂O和D₂O介质中的沉降测量,测得这种纯化的核心复合物的偏比容和S₂₀,w分别为0.70 cm³/g和14.5 S。根据平衡离心法估算,核心复合物的分子量为470000;平移摩擦系数与未溶剂化等效球体的摩擦系数之比经计算为1.9。用辛二酸二甲酯处理溶液中的游离促旋酶,可得到三种大致等量的交联产物,可鉴定为α₂、α₂β和α₂β₂。当用相同的交联剂处理促旋酶核心复合物时,70-80%的蛋白质转化为α₂β₂产物。这些结果表明,促旋酶-DNA核心复合物含有一段140个碱基对的DNA片段和一个α₂β₂四聚体蛋白。