S-Glutathionylation is a mechanism of signal transduction by which cells respond effectively and reversibly to redox inputs. The glutathionylation regulates most cellular pathways. It is involved in oxidative cellular response to insult by modulating the transcription factor Nrf2 and inducing the expression of antioxidant genes (ARE); it contributes to cell survival through nuclear translocation of NFkB and activation of survival genes, and to cell death by modulating the activity of caspase 3. It is involved in mitotic spindle formation during cell division by binding cytoskeletal proteins thus contributing to cell proliferation and differentiation. Glutathionylation also interfaces with the mechanism of phosphorylation by modulating several kinases (PKA, CK) and phosphatases (PP2A, PTEN), thus allowing a cross talk between the two processes of signal transduction. Also, skeletal RyR1 channels responsible of muscle excitation-contraction coupling appear to be sensitive to glutathionylation. Members of the ryanodine receptor super family, responsible for Ca(2) release from endoplasmic reticulum stores, contain sulfhydryl groups that function as a redox "switch", which either induces or inhibits Ca(2) release. Finally, but very importantly, glutathionylation of proteins may also act on cell metabolism by modulating enzymes involved in glycosylation, in the Krebs cycle and in mitochondrial oxidative phosphorylation. In this review, we propose a greater role for glutathionylation in cell biology: not only a cellular response to oxidative stress, but an elegant and sensitive mechanism able to respond even to subtle changes in redox balance in the different cellular compartments. Given the wide spectrum of redox-sensitive proteins, we discuss the possibility that different pathways light up by glutathionylation under various pathological conditions. The feature of reversibility of this process also makes it prone to develop targeted drug therapies and monitor the pharmacological effectiveness once identified the sensor proteins involved.