Novel insights into P450 BM3 interactions with FDA-approved antifungal azole drugs

Laura Jeffreys, Harshwardhan Poddar, Marina Golovanova, Colin Levy, Hazel Girvan, Kirsty Mclean, Michael W. Voice, David Leys, Andrew Munro

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Flavocytochrome P450 BM3 is a natural fusion protein constructed of cytochrome P450 and NADPH-cytochrome P450 reductase domains. P450 BM3 binds and oxidizes several mid- to long-chain fatty acids, typically hydroxylating these lipids at the ω-1, ω-2 and ω-3 positions. However, protein engineering has led to variants of this enzyme that are able to bind and oxidize diverse compounds, including steroids, terpenes and various human drugs. The wild-type P450 BM3 enzyme binds inefficiently to many azole antifungal drugs. However, we show that the BM3 A82F/F87V double mutant (DM) variant binds substantially tighter to numerous azole drugs than does the wild-type BM3, and that their binding occurs with more extensive heme spectral shifts indicative of complete binding of several azoles to the BM3 DM heme iron. We report here the first crystal structures of P450 BM3 bound to azole antifungal drugs – with the BM3 DM heme domain bound to the imidazole drugs clotrimazole and tioconazole, and to the triazole drugs fluconazole and voriconazole. This is the first report of any protein structure bound to the azole drug tioconazole, as well as the first example of voriconazole heme iron ligation through a pyrimidine nitrogen from its 5-fluoropyrimidine ring.
    Original languageEnglish
    Article number1577
    Pages (from-to)1-12
    Number of pages12
    JournalNature Scientific Reports
    Volume9
    Issue number1
    Early online date7 Feb 2019
    DOIs
    Publication statusPublished - 2019

    Keywords

    • cytochrome P450
    • P450 BM3
    • Mutations
    • Azole antifungal drugs
    • protein engineering
    • ligand binding
    • EPR spectroscopy

    Research Beacons, Institutes and Platforms

    • Biotechnology
    • Manchester Institute of Biotechnology

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