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Biophys J, February 2000, p. 1059-1069, Vol. 78, No. 2

Study by 23Na-NMR, 1H-NMR, and Ultraviolet Spectroscopy of the Thermal Stability of an 11-Basepair Oligonucleotide

Perrine Cahen,* Michel Luhmer,* Catherine Fontaine,dagger Claude Morat,dagger Jacques Reisse,* and Kristin Bartik*

 *Laboratoire de Chimie Organique E.P. (CP165/64), Université Libre de Bruxelles, 1050 Bruxelles, Belgium; and  dagger L.E.D.S.S., Université Joseph Fourier, 38041 Grenoble Cedex, France

23Na-NMR, 1H-NMR, and ultraviolet (UV) spectroscopy have been used to study the thermal stability of the double helix structure of an 11-basepair oligonucleotide. The denaturation curves obtained by 23Na-NMR and UV are analyzed using a two-state model. The melting temperature and Delta H0 obtained are identical within experimental error, suggesting that modifications in the ionic atmosphere, probed by 23Na-NMR, and the modifications in the basepair stacking, probed by UV, occur at the same temperature. Additional dynamical information on the denaturation process has been obtained by 1H-NMR: slow exchange is observed between the thymine methyl resonances, and the disappearance of imino protons shows that a single basepair opening does not contribute significantly to proton exchange.

Biophys J, February 2000, p. 1059-1069, Vol. 78, No. 2
© 2000 by the Biophysical Society   0006-3495/00/02/1059/11  $2.00




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