2′-alkynylnucleotides : a sequence- and spin label-flexible strategy for EPR Spectroscopy in DNA
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Electron paramagnetic resonance (EPR) spectroscopy is a powerful method to elucidate molecular structure through the measurement of distances between conformationally well-defined spin labels. Here we report a sequence-flexible approach to the synthesis of double spin-labeled DNA duplexes, where 2′-alkynylnucleosides are incorporated at terminal and internal positions on complementary strands. Post-DNA synthesis copper-catalyzed azide-alkyne cycloaddition (CuAAC) reactions with a variety of spin labels enable the use of double electron-electron resonance experiments to measure a number of distances on the duplex, affording a high level of detailed structural information.
Haugland , M M , El-Sagheer , A H , Porter , R J , Peña , J , Brown , T , Anderson , E A & Lovett , J E 2016 , ' 2′-alkynylnucleotides : a sequence- and spin label-flexible strategy for EPR Spectroscopy in DNA ' Journal of the American Chemical Society , vol 138 , no. 29 , pp. 9069-9072 . DOI: 10.1021/jacs.6b05421
Journal of the American Chemical Society
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TB and AHE-S thank the BBSRC (sLOLA grant BB/J001694/1). EAA thanks the EPSRC (EP/K005391/1, EP/M019195/1). JEL thanks the EPSRC (EP/LO22044/1), and the Royal Society for a University Research Fellowship and grant RF120645. We thank the Wellcome Trust for the Q-band EPR spectrometer (099149/Z/12/Z), the EPSRC National EPR Service for spectrometer time, and Dr Hassane El Mkami for technical assistance.
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