The conformational properties of a diastereomeric nucleoside-phosphorothioate pair is reported as reflected by different NMR parameters. Configurational assessment is based partly on the different NOE (nuclear overhauser enhancement) effects of the individual isomers and on the trends observed in other NMR parameters. Vicinal carbon-phosphorus couplings reflect the predominance of the epsilon(-) conformation for the Sp isomer and the epsilon(t) conformation for the Rp isomer. The effects of solvent and temperature on these vicinal couplings are described and the results are interpreted in terms of conformational shift towards the preferred epsilon(t) conformation.