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Its two-step synthesis — glutamate-cysteine ligase (GCL, rate-limiting, producing γ-glutamylcysteine at the cost of one ATP) followed by glutathione synthetase (adding glycine, consuming a second ATP) — produces a tripeptide whose unusual γ-glutamyl linkage between glutamate's γ-carboxylate and cysteine's amine renders it resistant to cellular peptidases, enabling accumulation to 0.5–15 mM concentrations in most cell types