Interference of ATP with the fluorescent probes YOYO-1 andYOYO-3 modifies the mechanical properties of intercalator-stained DNA confined in nanochannels.
Interference of ATP with the fluorescent probes YOYO-1 andYOYO-3 modifies the mechanical properties of intercalator-stained DNA confined in nanochannels.
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ATP干扰荧光探针yoyo-1 andyoyo-3会修饰局部染色的DNA的机械性能。
DOI：
10.1007/s00604-015-1495-7
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发表时间：
2015-06-01
期刊：
影响因子：
--
通讯作者：
Riehn R
中科院分区：
文献类型：
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作者：
Roushan M;Azad Z;Lim SF;Wang H;Riehn R
嵌入型荧光探针在DNA-蛋白质相互作用的研究中被广泛用于DNA的可视化。有些需要三磷酸腺苷（ATP）的存在。我们已经研究了DNA的机械性能染色的荧光嵌入染料YOYO-1和YOYO-3作为ATP浓度的函数（高达2 mM），通过拉伸单分子在纳米流体通道与通道横截面小到大约100×100 nm 2。ATP的存在使DNA的长度减少了11%。另一方面，如果用小沟结合探针4′，6-二脒基-2-苯基吲哚观察DNA，则发现可以忽略不计的影响。在纳米限制下延伸的明显下降归因于染料和ATP的相互作用，以及由此导致的YOYO-1从双螺旋中的排出。用YOYO-1染色的纳米通道拉伸的DNA（48.5kbp）对缓冲液中的ATP浓度敏感。使用横截面为80×80 nm 2的纳米通道拉伸DNA。
Intercalating fluorescent probes are widely used to visualize DNA in studies on DNA-protein interactions. Some require the presence of adenosine triphosphate (ATP). We have investigated the mechanical properties of DNA stained with the fluorescent intercalating dyes YOYO-1 and YOYO-3 as a function of ATP concentrations (up to 2 mM) by stretching single molecules in nanofluidic channels with a channel cross-section as small as roughly 100×100 nm2. The presence of ATP reduces the length of the DNA by up to 11 %. On the other hand, negligible effects are found if DNA is visualized with the minor groove-binding probe 4′,6-diamidino-2-phenylindole. The apparent drop in extension under nanoconfinement is attributed to an interaction of the dye and ATP, and the resulting expulsion of YOYO-1 from the double helix. Nanochannel-stretched DNA (48.5 kbp) stained with YOYO-1 is sensitive to ATP concentration in buffer. Nanochannels with a cross-section of 80×80 nm2 were used to stretch DNA.