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

DOI: 10.1038/s41598-018-37330-y DOI Help

Authors: Laura N. Jeffreys (Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), The University of Manchester) , Harshwardhan Poddar (Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), The University of Manchester) , Marina Golovanova (Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), The University of Manchester) , Colin Levy (Manchester Protein Structure Facility (MPSF), The University of Manchester) , Hazel M. Girvan (Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), The University of Manchester) , Kirsty J. Mclean (Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), The University of Manchester) , Michael W. Voice (Cypex Ltd) , David Leys (Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), The University of Manchester) , Andrew Munro (Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), The University of Manchester)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Scientific Reports , VOL 9

State: Published (Approved)
Published: February 2019
Diamond Proposal Number(s): 17773

Open Access Open Access

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.

Journal Keywords: Biophysical chemistry; Enzymes; Structural biology

Subject Areas: Biology and Bio-materials, Medicine


Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography

Documents:
s41598-018-37330-y.pdf