ZFIN Image: Enright et al., 2015, Fig. 3

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Figure Caption

Fig. 3 Cyp27c1 Is Sufficient to Convert Vitamin A1 into Vitamin A2

(A) HEK293 cells were transfected with a cyp27c1 expression construct or an empty vector and incubated with vitamin A1 for 24 hr, after which cells and media were harvested for HPLC. Cells transfected with empty vector did not produce vitamin A2, but cells transfected with cyp27c1 converted a substantial fraction of the vitamin A1 to vitamin A2.

(B) A cell-free assay system was used to assess the catalytic efficiency of Cyp27c1 for all-trans retinol, retinal, and retinoic acid. Purified Cyp27c1 was incubated with bovine adrenodoxin (Adx), NADPH-adrenodoxin reductase (ADR), and substrate (with concentrations ranging from 0.1 to 10 μM). The reaction was then initiated using an NADPH-generating system, run for 60 s, and quenched. HPLC was used to quantify the products in order to calculate kcat and Km. A high catalytic efficiency (kcat/Km) was observed for all three substrates. Error bars represent SEM. See also Figure S2.

Acknowledgments

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