T7 DNA polymerase reduced insulin at the same K m as thioredoxin, while the turnover number decreased. Recycling of the disulfide of thioredoxin subunit to its dithiol form was made by thioredoxin reductase. Incubation of T7 DNA polymerase with insulin decreases its ability to bind DNA and therefore inhibited polymerase and exonuclease activities. Thioredoxin reductase fully reversed this inhibition. Insulin did not induce dissociation of the T7 DNA polymerase subunits, which was tested by immunoadsorbent chromatography. No significant difference in single-stranded exonuclease compared to polymerase activity was seen in the flow through or the eluate, which had been expected if a dissociation of the subunits had occurred.
