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[采用¹H至³H交换法及电子显微镜法对溶液中聚(G)·聚(C)和聚(dG)·聚(dC)的分子间缔合进行研究]

[Study of the intermolecular association of poly(G).poly(c) and poly(dG).poly(dC) in solutions by methods of 1H to 3H exchange and electron microscopy].

作者信息

Lapiashvili G N, Lesnik E A, Kochkina I M, Maslova R N, Varshavskii Ia M

出版信息

Mol Biol (Mosk). 1979 Nov-Dec;13(6):1369-76.

PMID:547180
Abstract

The kinetic of 1H leads to 3H exchange between water and C(8)H-groups of the guanylic residues in poly(G) . poly(C) and poly(dG) . poly(dC) was investigated within the temperature range from 30 to 90 degrees in 0.5 M NaCl (pH 7.2). It was shown that the exchange in freshly dissolved preparations at temperatures lower than 50 degrees proceeds faster than that in the case of GMP. According to the ylide mechanism of the exchange reaction the observed acceleration of the exchange is considered as a consequence of associates formation in poly(G) . poly(c) and poly(dG) . poly(dC) solutions at temperatures lower than 50 degrees. Associates are stabilized by intermolecular hydrogen bonds in which N(7) atoms of guanylic residues take part. The increase of the temperature is accompanied by gradual disappearance of the exchange acceleration. The retardation of exchange, which is characteristic of most non-associated double-stranded polynucleotides and nucleic acids is observed at the temperatures above 60 degrees. The retardation points to thermal destruction of the associates at temperatures higher than 50 degrees. The associates which are characterized by ordered structure including several "side by side" arranged double-stranded molecules were observed by electron microscopy. The addition of EDTA to solutions as well as the increase of temperature leads to destruction of the associates whereas the addition of Mg2+ makes the associates more stable.

摘要

在0.5M NaCl(pH 7.2)溶液中,研究了温度范围为30至90摄氏度时,聚(G)·聚(C)和聚(dG)·聚(dC)中鸟苷酸残基的C(8)H基团与水之间1H到3H的交换动力学。结果表明,在低于50摄氏度的温度下,新溶解制剂中的交换比GMP中的交换进行得更快。根据交换反应的叶立德机理,观察到的交换加速被认为是聚(G)·聚(C)和聚(dG)·聚(dC)溶液在低于50摄氏度时形成缔合体的结果。缔合体通过鸟苷酸残基的N(7)原子参与的分子间氢键得以稳定。温度升高伴随着交换加速的逐渐消失。在高于60摄氏度的温度下,观察到了大多数非缔合双链多核苷酸和核酸特有的交换延迟。这种延迟表明在高于50摄氏度的温度下缔合体发生了热破坏。通过电子显微镜观察到了具有有序结构的缔合体,其中包括几个“并排”排列的双链分子。向溶液中添加EDTA以及升高温度都会导致缔合体的破坏,而添加Mg2+会使缔合体更稳定。

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