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El residuo His 467 del GSR a conotinuacin protona el anin GS para liberar la primera molcula de GSH

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Fifty l supernatant was mixed with 40 l 100 mM KHPO 4 (pH 7.0) and incubated for 5 min at 37 C, and then 10 l substrates (final 100 M) such as 3-mercaptopyruvate (3MP, Sigma-Aldrich), Na 2 S (Wako Pure Cheimcal Industries, Osaka, Japan) or a control H 2 O were added to incubate at 37 C for 15 min