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The second step, catalysed by glutathione synthetase, adds glycine to complete the tripeptide.² Of the three constituent amino acids, cysteine is by far the least abundant intracellularly, making its availability the primary bottleneck for glutathione production.² This is why cysteine is described as the rate-limiting substrate — even when glutamate and glycine are plentiful, glutathione synthesis cannot proceed without sufficient cysteine
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