AFM for analysis of structure and dynamics of DNA and protein-DNA complexes.

AFM for analysis of structure and dynamics of DNA and protein-DNA complexes.
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DOI:
10.1016/j.ymeth.2008.09.002
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发表时间:
2009-03
期刊:
影响因子:
4.8
通讯作者:
Shlyakhtenko, Luda S.
Shlyakhtenko, Luda S.
中科院分区:
生物学3区
文献类型:
--
作者:
Lyubchenko, Yuri L.;Shlyakhtenko, Luda S.

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本文介绍了原子力显微镜利用表面化学方法研究DNA和蛋白质-DNA复合物的结构和动力学的协议。提供了使用硅烷和杂氮硅三环制备功能化表面和AFM探针的必要细节,包括合成杂氮硅三环的方案。局部和全局构象DNA的研究方法,主要集中在水溶液中的DNA的延时成像的霍利迪路口,包括分支迁移的动力学研究说明。以核小体动力学分析为例,说明了延时原子力显微镜在蛋白质-DNA复合物动力学研究中的应用。力谱是原子力显微镜的一种形式,在生物医学研究的各个领域都具有重要的意义。通过对突触SfiI-DNA复合物动力学分析的数据,说明了AFM力谱方法用于研究特定的蛋白质-DNA复合物。必要时,将向相应的示例添加其他细节。
This paper describes protocols for studies of structure and dynamics of DNA and protein-DNA complexes with AFM utilizing the surface chemistry approach. The necessary specifics for the preparation of functionalized surfaces and AFM probes with the use of silanes and silatranes, including the protocols for synthesis of silatranes are provided. The methodology of studies of local and global conformations DNA with the major focus on the time-lapse imaging of DNA in aqueous solutions is illustrated by the study of dynamics of Holliday junctions including branch migration. The analysis of nucleosome dynamics is selected as an example to illustrate the application of the time-lapse AFM to studies of dynamics of protein-DNA complexes. The force spectroscopy is the modality of AFM with a great importance to various fields of biomedical studies. The AFM force spectroscopy approach for studies of specific protein-DNA complexes is illustrated by the data on analysis of dynamics of synaptic SfiI-DNA complexes. When necessary, additional specifics are added to the corresponding example.
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