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In addition, deletion of the GSH1 geneencoding -glutamylcysteine synthetase (-GCS), the enzyme catalysing the first step in glutathione synthesiswas shown to suppress an Ero1 conditional mutant, again indicating that GSH provides an additional load on Ero1 activity (Frand & Kaiser, 1998)

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The membranes were washed and incubated at room temperature for 1 h with Amersham ECL horseradish peroxidase-conjugated immunoglobulin G F(ab)2 fragment antibody (1:5,000 dilution