Figure 4 | Scientific Reports

Figure 4

From: Development of a Synthetic 3-ketosteroid Δ1-dehydrogenase for the Generation of a Novel Catabolic Pathway Enabling Cholesterol Degradation in Human Cells

Figure 4

Δ1-KstDR expression in Hep3B cells enables novel catabolic activity. Representative HPLC chromatograms (λ 245 nm) showing PD (tr = 13.8 min) in control Hep3B cells (a) and Hep3B expressing Δ1-KstD (b) after 24 showing the accumulation of PDD (λmax: 247 nm; tr = 10.0 min) only in the Hep3B cells expressing Δ1-KstD. Additional data and images showing [14C] trace detected by the in-line scintillation detector corresponding to the chromatograms are provided in Figs S20 and S21. For all experiments, Hep3B controls (left) or Hep3B Δ1-KstD cells (right) were incubated with 15.7 μg (10 μM) PD spiked with 100 nCi C4-[14C] labeled PD (tr = 13.8 min). (c) Structures of substrates, products, and a diagram of the reaction summary following catabolism of PD generating PDD and catabolism of 9-hydroxypregn-4-ene-3,20-dione (9-OHPD) generating 3-hydroxy-9,10-secopregn-1,3,5(10)-triene-9,20-dione (3-HSP).

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