Cigarette smoke causes irreversible oxidations in lungs, but its impact on reversible and physiologically relevant redox-dependent protein modifications remains to be investigated. Here the effect of cigarette smoke exposure in mice was investigated on the covalent binding of glutathione to protein thiols, known as S-glutathionylation (PSSG), which can be reversed by glutaredoxins (Grx). Also, protein S-nitrosylation (PSNO) which is the modification of protein thiols by NO and which is reversed by the enzyme alcohol dehydrogenase (ADH) 5 was examined. Both PSSG and PSNO levels in lung tissue were markedly decreased after 4 weeks of cigarette smoke exposure. This coincided with attenuated protein free thiol levels and increased protein carbonylation. The expression of NOX4, DHE sensitive oxidant production and iNOS levels were induced by smoke, whereas Grx1 mRNA expression and activity were attenuated. Free GSH levels, protein expression and activity of ADH5 were unaffected by smoke. Taken together, smoke exposure decreases reversible cysteine oxidations PSSG and PSNO and enhances protein carbonylation. These alterations are not associated with differences in some of the regulatory enzymes, but are likely the result of oxidative stress.