[6] This enzyme utilizes and stabilizes an acylphosphate intermediate to later perform a favorable nucleophilic attack of glycine.

[citation needed] Human and yeast glutathione synthetases are homodimers, meaning they are composed of two identical subunits of itself non-covalently bound to each other.

Sulfate ion serves as a replacement for inorganic phosphate once the acylphosphate intermediate is formed inside the active site.

In Arabidopsis thaliana, low levels of glutathione synthetase have resulted in increased vulnerability to stressors such as heavy metals, toxic organic chemicals, and oxidative stress.

[15] The presence of a thiol functional group allows its product GSH to serve both as an effective oxidizing and reducing agent in numerous biological scenarios.

Additionally, the thiol side chain of cysteines serve as potent nucleophiles and react with oxidants and electrophilic species that would otherwise cause damage to the cell.

[19] Because of its antioxidant role, GSS mostly produce GSH inside the cytoplasm of liver cells and imported to mitochondria where detoxification occurs.

[20] GSH is also essential for the activation of the immune system to generate robust defense mechanisms against invading pathogens.

Moderately and severely affected individuals have enzymes with dysfunctional catalytic sites, rendering it unable to participate in detoxification reactions.

Physiological symptoms include metabolic acidosis, neurological defects, and increased susceptibility to pathogenic infections.

In order to treat metabolic acidosis, severely affected patients are given large amounts of bicarbonate and antioxidants such as vitamin E and vitamin C.[24] In mild cases, ascorbate and N-acetylcysteine have been shown to increase glutathione levels and increase erythrocyte production.