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. 2023 May 18;299(7):104841. doi: 10.1016/j.jbc.2023.104841

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© 2023 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

LC-UV assay of conversion of dihydrolanosterol oxidation to dihydro FF-MAS. UPLC traces of a reaction of 10 μM dihydrolanosterol with 0.05 μM P450 51A1 for 5 min. The products were extracted and analyzed by UPLC. A, A₁₉₂; B, A₂₅₀ measurements. Peaks for the residual lanosterol after the reaction (A, t_R 14.1 min) and the demethylated product, dihydro FF-MAS (B, t_R 9.9 min), are shown. The intermediates (14α-alcohol and 14α-aldehyde) were not detected in steady-state experiments but absorb maximally at 192 nm and 230 nm, respectively. The A₂₅₀ character of the dihydro FF-MAS product is the result of the additional degree of unsaturation of the diene. FF-MAS, follicular fluid meiosis-activating sterol ((4β,5α)-4,4-dimethyl-cholesta-8,14,24-trien-3-ol); UPLC, ultra-performance liquid chromatography.