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Methods Cloning and purification Three hTrxR1 constructs were cloned into the pET28a(+) expression vector (Novagen) using Nde I and Hin dIII restriction sites: a double hTrxR1 mutant where Cys497 and Sec498 were replaced by Ser and Cys, respectively (hTrxR1 C497S,U498C ), as well as a hTrxR1 mutant where Sec498 was replaced by Ser (hTrxR1 U498S ), and a truncated form of hTrxR1 where a stop codon was introduced after Cys497 (hTrxR1 C497_trunc )