通过闪光猝灭技术与金属氧络合物的直接氧化作用对 RNA 中鸟嘌呤的氧化性质进行研究。
Nature of guanine oxidation in RNA via the flash-quench technique versus direct oxidation by a metal oxo complex.
机构信息
Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina 27599-3290, USA.
出版信息
Inorg Chem. 2010 Feb 1;49(3):786-95. doi: 10.1021/ic9008619.
Oxidation of RNA can be effected by two different techniques: a photochemical, electron-transfer method termed "flash-quench" and direct oxidation by metal oxo complexes. The flash-quench method produces selective oxidation using a metal photosensitizer, tris(bipyridyl)ruthenium(III) trichloride (Ru(bpy)(3)(3+)), and quencher, pentaamminechlorocobalt(III) chloride (Co(NH(3))(5)Cl(2+)). We have optimized the flash-quench technique for the following RNAs: tRNA(Phe), human ferritin iron-responsive element (IRE), and a mutated human ferritin IRE. We have also employed a chemical footprinting technique involving the oxoruthenium(IV) complex (Ru(tpy)(bpy)O(2+) (tpy = 2,2',2''-terpyridine; bpy = 2,2'-bipyridine)) to oxidize guanine. Comparison of the two methods shows that the flash-quench technique provides a visualization of nucleotide accessibility for a static conformation of RNA while the Ru(tpy)(bpy)O(2+) complex selectively oxidizes labile guanines and gives a visualization of a composite of multiple conformations of the RNA structure.
RNA 的氧化可以通过两种不同的技术来实现：一种是光化学的、电子转移方法，称为“闪光猝灭”，另一种是金属氧络合物的直接氧化。闪光猝灭法使用金属光敏剂三（联吡啶）钌（III）三氯化物（Ru（bpy）（3）（3+））和猝灭剂五氨氯化钴（III）（Co（NH3）（5）Cl2+）产生选择性氧化。我们已经优化了以下 RNA 的闪光猝灭技术：tRNA（Phe）、人铁蛋白反应元件（IRE）和突变的人铁蛋白 IRE。我们还采用了一种涉及氧钌（IV）配合物（Ru（tpy）（bpy）O2+（tpy=2,2',2''-三联吡啶；bpy=2,2'-联吡啶）的化学足迹技术来氧化鸟嘌呤。两种方法的比较表明，闪光猝灭技术提供了 RNA 静态构象中核苷酸可及性的可视化，而 Ru（tpy）（bpy）O2+配合物则选择性地氧化不稳定的鸟嘌呤，并提供 RNA 结构的多个构象的组合的可视化。
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