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Cleavage of supercoiled DNA by deoxyribonuclease I in solution and at the electrode surface

机译：溶液中和电极表面的脱氧核糖核酸酶I切割超螺旋DNA

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Cleavage of supercoiled DNA by deoxyribonuclease I (DNase I) in solution and at the surface of the mercury electrode was studied by means of AC voltammetry. This technique produces peak 3 which is produced only by DNAs containing free ends (such as linear double-stranded and single-stranded DNAs and open circular DNAs) but not by covalently closed circular (ccc) DNAs. Formation of a single interruption of the sugar-phosphate backbone in the ccc supercoiled (sc) DNA results in formation of peak 3. Peak 1 is produced by both ccc DNA molecules as well as by DNAs containing free ends; changes in height of this peak occur due to DNA cleavage. We show that the kinetics of the cleavage of DNA in solution and at the electrode surface substantially differ suggesting restricted accessibility of the surface-confined DNA for the interaction with the enzyme. Cleavage of the immobilized DNA is remarkably influenced by the potential of the electrode surface. At positively charged surface the enzymatic reaction is inhibited in its initial stage while moderately negative charges stimulate the cleavage of the immobilized DNA by DNase I. [References: 31]
机译：通过交流伏安法研究了溶液中和汞电极表面的脱氧核糖核酸酶I（DNase I）对超螺旋DNA的裂解。此技术产生的峰3仅由包含自由端的DNA（例如线性双链和单链DNA和开放的环状DNA）产生，而不由共价闭合的环状（ccc）DNA产生。 ccc超螺旋（sc）DNA中糖磷酸主链的单次中断形成导致峰3的形成。峰1由ccc DNA分子和包含自由端的DNA产生;该峰的高度变化是由于DNA裂解而发生的。我们表明，在溶液中和在电极表面处的DNA切割动力学基本不同，这表明与酶相互作用的表面受限DNA的可及性受到限制。固定的DNA的切割受电极表面电势的显着影响。在带正电荷的表面，酶反应在其初始阶段受到抑制，而适度的负电荷则刺激DNase I对固定的DNA的切割。[参考文献：31]

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