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Comparison of velocity versus substrate plots for wild type AKR1D1 and the P133R mutant using Δ4-cholesten-7α-ol-3-one as substrate. ■, AKR1D1; ▴, P133R. Assays were performed using standard fluorimetric assay conditions. For the wild type enzyme, the experiment was replicated twice, and for the P133R mutant, the experiment was replicated once. Measurements were done in duplicate, and variation was less than 10%. One representative experiment is shown.
