Potentiometric determinations of the two successive acidities of nicotinic and isonicotinic acids (AH2+) have been carried out in aqueous dimethyl sulfoxide mixtures containing up to 95% Me2SO by volume. In both systems, the results reveal that the addition of Me2SO induces a proton transfer from the pyridinium ring to the carboxylate group, the tautomeric equilibrium between the neutral forms of the two acids being displaced toward the zwitterionic form (AH±) in aqueous solution, but toward the molecular form (AH°) in Me2SO. An analysis of the data by means of Hammett relationships previously established for benzoic acids over the whole range of H2O/Me2SO mixtures allowed the four microscopic acidity constants as well as the tautomeric equilibrium constant KT pertaining to the complete ionization scheme of the two acids to be determined. At 20 °C, there are equal populations of the tautomeric AH° and AH± species in the mixtures containing 38 and 47% Me2SO for the nicotinic and isonicotinic systems, respectively. Hammett relationships describing the ionization behaviour of a number of substituted pyridinium cations in H2O/Me2SO mixtures are also discussed. Possible reasons accounting for the peculiar effects exerted by the NH2, CONH2, and COOH substituents on the process are suggested. Key words: nicotinic and isonicotinic acids, substituted pyridines, acidities, tautomeric equilibrium, protonation sites, water – dimethyl sulfoxide mixtures.