Flaender M, Sicoli G, Fontecave Th, Mathis G, Saint-Pierre C, Boulard Y, Gambarelli S, Gasparutto D
Laboratoire des Lésions des Acides Nucléiques, SCIB/UMR E3 CEA-UJF, INAC, CEA-Grenoble, F-38054 Grenoble Cedex 9, France.
Nucleic Acids Symp Ser (Oxf). 2008(52):147-8. doi: 10.1093/nass/nrn075.
A new approach is described for the insertion of nitroxide spin-labels at specific positions within DNA oligomers. The latter bioconjugaison strategy is based on a click chemistry 1,3-dipolar cycloaddition between a spin-labeling reagent, namely the 4-azido-TEMPO, and alkyne modified uridine-containing oligonucleotides. This highly efficient labeling method was applied for site-specific incorporation of two TEMPO units within a set of double-stranded DNA constructs. Then the determination of the inter-nitroxide distances was achieved by using a four-pulses DEER technique that successfully validates the new site-directed spin labeling strategy.
本文描述了一种在DNA寡聚物特定位置插入氮氧化物自旋标记的新方法。后一种生物共轭策略基于自旋标记试剂（即4-叠氮基-TEMPO）与炔烃修饰的含尿苷寡核苷酸之间的点击化学1,3-偶极环加成反应。这种高效的标记方法被用于在一组双链DNA构建体中位点特异性地掺入两个TEMPO单元。然后，通过使用四脉冲双电子-电子共振（DEER）技术测定氮氧化物之间的距离，该技术成功验证了新的位点定向自旋标记策略。
