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Its two-step synthesis glutamate-cysteine ligase (GCL, rate-limiting, producing -glutamylcysteine at the cost of one ATP) followed by glutathione synthetase (adding glycine, consuming a second ATP) produces a tripeptide whose unusual -glutamyl linkage between glutamate's -carboxylate and cysteine's amine renders it resistant to cellular peptidases, enabling accumulation to 0.515 mM concentrations in most cell types

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