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光可逆凝聚过程中DNA分子的构象与动力学

Conformation and dynamics of DNA molecules during photoreversible condensation.

作者信息

Le Ny Anne-Laure M, Lee C Ted

机构信息

Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90089-1211, USA.

出版信息

Biophys Chem. 2009 Jun;142(1-3):76-83. doi: 10.1016/j.bpc.2009.03.010. Epub 2009 Mar 31.

Abstract

Direct observation of the mechanism and dynamics of photo-initiated DNA compaction and re-expansion using a light-responsive cationic surfactant has been achieved with fluorescence microscopy. The surfactant undergoes a reversible photoisomerization upon exposure to visible (trans isomer, relatively hydrophobic) or UV (cis isomer, relatively hydrophilic) light. Thus, surfactant binding to DNA and the DNA condensation that result can both be initiated and controlled with light illumination. The inherent kinetics of DNA conformational changes, directly visualized following the in situ light "trigger" of surfactant photoisomerization, are found to occur at rates of approximately 9 microm/s or 240 kbp/s, at or near rates that can be achieved in natural processes. Furthermore, observation of photo-initiated DNA compaction, free of the effects of shear or mixing, provides evidence of a condensation mechanism that nucleates at the ends of the macromolecule. Ethidium bromide displacement studies, employed to gain insight on the mode of interaction between the photo-surfactant and DNA, also reveal the importance of both electrostatic and hydrophobic forces in surfactant binding and DNA condensation.

摘要

利用荧光显微镜已实现对使用光响应性阳离子表面活性剂的光引发DNA压缩和再膨胀的机制及动力学的直接观察。该表面活性剂在暴露于可见光(反式异构体,相对疏水)或紫外线(顺式异构体,相对亲水)时会发生可逆的光异构化。因此,表面活性剂与DNA的结合以及由此导致的DNA凝聚都可以通过光照引发和控制。在原位光“触发”表面活性剂光异构化后直接观察到的DNA构象变化的固有动力学,发现其发生速率约为9微米/秒或240千碱基对/秒,处于或接近自然过程中可实现的速率。此外,对不受剪切或混合影响的光引发DNA压缩的观察,为在大分子末端成核的凝聚机制提供了证据。用于深入了解光表面活性剂与DNA之间相互作用模式的溴化乙锭置换研究,也揭示了静电和疏水作用力在表面活性剂结合和DNA凝聚中的重要性。

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