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The absorption spectrum of CYP125RHA1 in the oxidized state as isolated (solid line) and incubated with 10 μM cholesterol in oxidized (dashed line) and reduced (dotted line) states. The inset shows the reduced CO-difference spectrum of the enzyme incubated with 10 μM cholesterol. The sample contained 2.9 μM purified CYP125RHA1, 0.1 M potassium phosphate buffer, pH 7.0, 25°C; cholesterol was added from a 1 mM stock solubilized in 10% 2-hydroxypropyl-β-cyclodextrin.
