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[通过原子力显微镜研究支撑底物对DNA及DNA-三缬氨酸复合物结构的影响]

[Effect of supporting substrates on the structure of DNA and DNA-trivaline complexes studied by atomic force microscopy].

作者信息

Klinov D V, Martynkina L P, Iurchenko V Iu, Demin V V, Strel'tsov S A, Gerasimov Iu A, Vengerov Iu Iu

机构信息

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997 Russia.

出版信息

Bioorg Khim. 2003 Jul-Aug;29(4):397-402. doi: 10.1023/a:1024901318099.

Abstract

Linear DNA, circular DNA, and circular DNA complexes with trivaline (TV), a synthetic oligopeptide, were imaged by atomic force microscopy (AFM) using mica as a conventional supporting substrate and modified highly ordered pyrolytic graphite (HOPG) as an alternative substrate. A method of modifying the HOPG surface was developed that enabled the adsorption of DNA and DNA-TV complexes onto this surface. On mica, both purified DNA and DNA-TV complexes were shown to undergo significant structural distortions: DNA molecules decrease in height and DNA-TP displays substantial changes in the shape of its circular compact structures. Use of the HOPG support helps preserve the structural integrity of the complexes and increase the measured height of DNA molecules up to 2 nm. AFM with the HOPG support was shown to efficiently reveal the particular points of the complexes where, according to known models of their organization, a great number of bent DNA fibers meet. These results provide additional information on DNA organization in its complexes with TV and are also of methodological interest, since the use of the modified HOPG may widen the possibilities of AFM in studying DNA and its complexes with various ligands.

摘要

使用云母作为传统支撑基底以及改性的高度有序热解石墨(HOPG)作为替代基底,通过原子力显微镜(AFM)对线性DNA、环状DNA以及环状DNA与三缬氨酸(TV,一种合成寡肽)的复合物进行了成像。开发了一种改性HOPG表面的方法,该方法能够使DNA和DNA - TV复合物吸附到该表面上。在云母上,纯化的DNA和DNA - TV复合物均显示出明显的结构扭曲:DNA分子高度降低,DNA - TV其环状紧密结构的形状发生显著变化。使用HOPG支撑有助于保持复合物的结构完整性,并使DNA分子的测量高度增加至2 nm。结果表明,带有HOPG支撑的AFM能够有效地揭示复合物的特定点,根据其已知的组织模型,在这些特定点有大量弯曲的DNA纤维相交。这些结果提供了有关DNA与TV形成复合物时其组织形式的额外信息,并且在方法学上也具有重要意义,因为使用改性HOPG可能会拓宽AFM在研究DNA及其与各种配体形成的复合物方面的可能性。

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