Selectively 13C-enriched DNA: 13C and 1H assignments of a triple helix by two-dimensional relayed HMQC experiments |
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Authors: | Olivier Bornet Gérard Lancelot Luc Chanteloup Nguyen T Thuong Jean-Marie Beau |
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Institution: | (1) Centre de Biophysique Moléculaire, CNRS, 1A Avenue de la Recherche Scientifique, F-45071 Orléans Cedex 2, France;(2) Laboratoire de Biochimie Structurale associé au CNRS, Université d'Orléans, BP 6759, F-45067 Orléans Cedex 2, France |
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Abstract: | Summary We present NMR studies of an intramolecular triple helix, the three strands of which have been linked by a hexaethylene glycol chain. To overcome the generally encountered difficulties of assignment in the homopyrimidine strands, the carbon C1 of the pyrimidines were selectively 13C-enriched. Assignments of the aromatic and sugar protons were obtained from NOESY-HMQC and TOCSY-HMQC spectra. We show that the recognition of a DNA duplex by a third strand via triplex formation is easily carried out in solution by observing the changes of the 1H1–13C1 connectivities as a function of pH. Furthermore, the conformation of the sugars has been found to be C2-endo, on the basis of the coupling constant values directly measured in an HSQC spectrum. |
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Keywords: | 13C-labeled oligonucleotides Triple helix |
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