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双链和单链DNA与精胺、亚精胺、腐胺、二氨基丙烷、N1-和N8-乙酰亚精胺以及镁相互作用的缔合常数:通过热变性曲线展宽分析测定

Association constants for the interaction of double-stranded and single-stranded DNA with spermine, spermidine, putrescine, diaminopropane, N1- and N8-acetylspermidine, and magnesium: determination from analysis of the broadening of thermal denaturation curves.

作者信息

Morgan J E, Blankenship J W, Matthews H R

出版信息

Arch Biochem Biophys. 1986 Apr;246(1):225-32. doi: 10.1016/0003-9861(86)90467-4.

Abstract

The effect of Mg2+, putrescine, diaminopropane, N1-acetylspermidine, N8-acetylspermidine, spermidine, and spermine on the thermal denaturation of calf thymus DNA was investigated. As in a previous study with magnesium [W.F. Dove and N. Davidson, (1962) J. Mol. Biol. 5, 467-478], these ligands were found to raise the thermal denaturation temperature of the DNA and to broaden the thermal denaturation curve dramatically at the point where 10 to 20% of the DNA charge had been neutralized. At higher levels of charge neutralization the curves became sharper again. This behavior was due to differential binding of the ligands to single- and double-stranded DNA. The broadening was used to determine the ratio of the association constants of each ligand to the two forms of DNA using either an independent sites model of binding or an excluded sites model. The results show that the primary mode of binding of the ligands to DNA is electrostatic but that important secondary, nonelectrostatic, effects are also present.

摘要

研究了Mg2+、腐胺、二氨基丙烷、N1-乙酰亚精胺、N8-乙酰亚精胺、亚精胺和精胺对小牛胸腺DNA热变性的影响。正如之前关于镁的研究[W.F. Dove和N. Davidson,(1962年)《分子生物学杂志》5,467 - 478]所示,发现这些配体可提高DNA的热变性温度,并在DNA电荷的10%至20%被中和时显著拓宽热变性曲线。在更高的电荷中和水平下,曲线再次变陡。这种行为是由于配体与单链和双链DNA的差异结合所致。利用结合的独立位点模型或排阻位点模型,通过拓宽曲线来确定每种配体与两种形式DNA的缔合常数之比。结果表明,配体与DNA结合的主要模式是静电作用,但也存在重要的次要的非静电作用。

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