Twisted cyanines: a non-planar fluorogenic dye with superior photostability and its use in a protein-based fluoromodule.
Twisted cyanines: a non-planar fluorogenic dye with superior photostability and its use in a protein-based fluoromodule.
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扭曲花青：一种具有优异光稳定性的非平面荧光染料及其在基于蛋白质的荧光模块中的应用。
DOI：
10.1021/ja308629w
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发表时间：
2013-01-09
影响因子：
15
通讯作者：
Armitage, Bruce A.
中科院分区：
文献类型：
--
作者：
Shank, Nathaniel I.;Pham, Ha H.;Waggoner, Alan S.;Armitage, Bruce A.
菁染料噻唑橙(TO)是一种广为人知的DNA和RNA荧光染料，但这一特性使其无法用作非核酸生物分子的细胞内荧光探针。此外，就像许多花菁一样，这种染料的光稳定性较低。在这里，我们报道了一种桥式取代的TO的合成，命名为α-CN-TO，其中TO的中心亚甲基氢被一个吸电子的氰基取代，这有望降低染料对单重态氧介导的降解的敏感性。X-射线晶体结构表明，α-CN-TO发生了剧烈的平面外扭曲，而TO则以平面构象结晶。α-CN-TO保留了母体染料TO在粘性甘油/水溶剂中的荧光行为，但直接照射和间接漂白研究表明，α-CN-TO对可见光和单线态氧基本上是惰性的。此外，α-CN-TO的扭曲构象减轻了DNADNA和预先选择的TO结合蛋白的非特异性结合和荧光激活，并在HeLa细胞培养中显示出低背景荧光。然后使用α-CN-TO来选择一种新的蛋白质，该蛋白质可以结合并激活染料中的荧光。新的α-CN-TO/蛋白质荧光模块表现出比类似/蛋白质荧光模块更好的光稳定性。这些性质表明，α-CN-TO将是一种有用的荧光染料，与特定的核糖核酸和蛋白质结合伙伴结合，在体外和细胞基础上都有应用。更广泛地说，促进非平面构象的结构特征可以提供一种有效的方法来减少阳离子染料与核酸和其他生物分子的非特异性结合。
The cyanine dye thiazole orange (TO) is a well-known fluorogenic stain for DNA and RNA, but this property precludes its use as an intracellular fluorescent probe for non-nucleic acid biomolecules. Further, as is the case with many cyanines, the dye suffers from low photostability. Here we report the synthesis of a bridge-substituted version of TO named α-CN-TO, where the central methine hydrogen of TO is replaced by an electron withdrawing cyano group, which was expected to decrease the susceptibility of the dye toward singlet oxygen-mediated degradation. An X-ray crystal structure shows that α-CN-TO is twisted drastically out of plane, in contrast to TO, which crystallizes in the planar conformation. α-CN-TO retains the fluorogenic behavior of the parent dye TO in viscous glycerol/water solvent, but direct irradiation and indirect bleaching studies showed that α-CN-TO is essentially inert to visible light and singlet oxygen. In addition, the twisted conformation of α-CN-TO mitigates non-specific binding and fluorescence activation by DNA and a previously selected TO-binding protein and exhibits low background fluorescence in HeLa cell culture. α-CN-TO was then used to select a new protein that binds and activates fluorescence from the dye. The new α-CN-TO/protein fluoromodule exhibits superior photostability to an analogous TO/protein fluoromodule. These properties indicate that α-CN-TO will be a useful fluorogenic dye in combination with specific RNA and protein binding partners for both in vitro and cell-based applications. More broadly, structural features that promote nonplanar conformations can provide an effective method for reducing nonspecific binding of cationic dyes to nucleic acids and other biomolecules.
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影响因子：
4.7
作者：
Fitzpatrick JA;Yan Q;Sieber JJ;Dyba M;Schwarz U;Szent-Gyorgyi C;Woolford CA;Berget PB;Waggoner AS;Bruchez MP
通讯作者：
Bruchez MP
影响因子：
2.6
作者：
Dash, S.;Panigrahi, M.;Mishra, B. K.
通讯作者：
Mishra, B. K.
影响因子：
15
作者：
AUBRY, JM
通讯作者：
AUBRY, JM
影响因子：
5.2
作者：
Constantin, Tudor P.;Silva, Gloria L.;Armitage, Bruce A.
通讯作者：
Armitage, Bruce A.
影响因子：
56.9
作者：
Debler, Erik W.;Kaufmann, Gunnar F.;Lerner, Richard A.
通讯作者：
Lerner, Richard A.