The periparturient period is characterized by increased oxidative stress status. Glutathione is one of the major intracellular antioxidants and is important for alleviating oxidative stress. Nuclear factor (erythroid-derived 2)-like factor 2 (NFE2L2) is a transcription factor that induces antioxidant-responsive genes. The hypothesis was that enhanced methionine supply has a positive effect on molecular pathways associated with glutathione synthesis and activation of NFE2L2 in adipose tissue. Sixty multiparous Holstein cows were used in a block design and assigned to a control diet or the control plus ethylcellulose rumen-protected methionine (MET; Mepron, Evonik Nutrition & Care GmbH, Germany). Mepron was fed from −28 to 30 d relative to parturition at 0.09% and 0.10% of diet dry matter during the prepartum and postpartum period, respectively. Adipose tissue obtained on d −10, 10, and 30 relative to calving date was used for RT-PCR analysis. Blood was sampled at −14, 7, and 30 d relative to calving date. The statistical model included the random effect of block and fixed effect of treatment, time and its interaction. Compared with control, MET cows had lower plasma concentrations of reactive oxygen metabolites (P = 0.08) and higher concentrations of methionine (P = 0.09) and glutathione (P = 0.02). Among enzymes associated with the methionine cycle and transsulfuration, expression of cystathionine-β-synthase in adipose tissue was upregulated in MET cows (P = 0.06). Expression of glutamate-cysteine ligase, glutathione reductase, and transaldolase 1 in adipose tissue also was upregulated in MET cows (P < 0.05), suggesting a positive effect of enhanced methionine supply on intracellular reduced glutathione availability. The upregulation of NFE2L2 in MET cows also is suggestive of a link between adipose tissue methionine metabolism, glutathione synthesis, and the antioxidant response (P = 0.06). No interactions with time were observed for gene expression in adipose tissue. Overall, the data indicate that enhanced methionine supply during the periparturient period can benefit the availability of reduced glutathione and activation of antioxidant-responsive genes in adipose tissue.