Thioredoxin, glutaredoxin, and thioredoxin reductase from cultured HeLa cells.

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Abstract
Thioredoxin and glutaredoxin may be important in regulating cell metabolism by mediating interchanges between sulfhydryl and disulfide groups. Components of the thioredoxin/glutaredoxin system from cultured HeLa cells have been partially purified and characterized by using Escherichia coli adenosine 3'-phosphate 5'-phosphosulfate reductase, a thioredoxin/glutaredoxin-dependent enzyme on the pathway of sulfate reduction, as an assay system. In HeLa cells, a NADPH-thioredoxin reductase and three heat-labile proteins (designated PI, PII, and PIII) that have thioredoxin- or glutaredoxin-like properties are found. Both PI and PIII have molecular masses of approximately 12,000 daltons and are readily reduced by their homologous HeLa thioredoxin reductase. However, only PI can be reduced efficiently by the glutathione system and neither PI nor PIII has inherent glutathione-disulfide oxidoreductase activity. PII has a molecular mass of greater than 30,000 daltons and appears to be associated with a reductase activity. The HeLa NADPH-thioredoxin reductase has been purified to near homogeneity and found to be a 116,000-dalton flavoprotein composed of two 58,000-dalton subunits. The HeLa enzyme has low species and substrate specificity and can reduce HeLa PI and PIII, E. coli thioredoxin and glutaredoxin, and the disulfide bond in 5,5'-dithiobis(2-nitrobenzoic acid). The exact in vivo roles of the HeLa thioredoxin/glutaredoxin system remain to be determined.