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dc.contributor.authorBowen, Alice M.
dc.contributor.authorJohnson, Eachan O. D.
dc.contributor.authorMercuri, Francesco
dc.contributor.authorHoskins, Nicola J.
dc.contributor.authorQiao, Ruihong
dc.contributor.authorMcCullagh, James S. O.
dc.contributor.authorLovett, Janet E.
dc.contributor.authorBell, Stephen G.
dc.contributor.authorZhou, Weihong
dc.contributor.authorTimmel, Christiane R.
dc.contributor.authorWong, Luet Lok
dc.contributor.authorHarmer, Jeffrey R.
dc.identifier.citationBowen , A M , Johnson , E O D , Mercuri , F , Hoskins , N J , Qiao , R , McCullagh , J S O , Lovett , J E , Bell , S G , Zhou , W , Timmel , C R , Wong , L L & Harmer , J R 2018 , ' A structural model of a P450-ferredoxin complex from orientation-selective double electron-electron resonance spectroscopy ' , Journal of the American Chemical Society , vol. 140 , no. 7 , pp. 2514-2527 .
dc.identifier.otherPURE: 252001352
dc.identifier.otherPURE UUID: 430d6bcc-8c18-45fd-9f50-cade6f1c013e
dc.identifier.otherScopus: 85042365238
dc.identifier.otherORCID: /0000-0002-3561-450X/work/45525136
dc.identifier.otherWOS: 000426143800024
dc.descriptionThis research was supported by the Engineering & Physical Sciences Research Council (EPSRC) and the Biotechnology & Biological Sciences Research Council (BBSRC), UK (EP/D048559). AMB and EOJD were supported by graduate studentships from the BBSRC (BB/F01709X/1) and NJH and JEL were supported by graduate studentships from the EPSRC, and JEL after her DPhil by EP/D048559. AMB gratefully acknowledges her current fellowship support from the Royal Society and EPSRC for a Dorothy Hodgkin Fellowship (DH160004). JRH acknowledges support from the ARC (FT120100421) and the Centre for Advanced Imaging, The University of Queensland.en
dc.description.abstractCytochrome P450 (CYP) monooxygenases catalyze the oxidation of chemically inert carbon-hydrogen bonds in diverse endogenous and exogenous organic compounds by atmospheric oxygen. This C–H bond oxy-functionalization activity has huge potential in biotechnological applications. Class I CYPs receive the two electrons required for oxygen activation from NAD(P)H via a ferredoxin reductase and ferredoxin. The interaction of Class I CYPs with their cognate ferredoxin is specific. In order to reconstitute the activity of diverse CYPs, structural characterization of CYP-ferredoxin complexes is necessary, but little structural information is available. Here we report a structural model of such a complex (CYP199A2-HaPux) in frozen solution derived from distance and orientation restraints gathered by the EPR technique of orientation-selective double electron-electron resonance (os-DEER). The long-lived oscillations in the os-DEER spectra were well modeled by a single orientation of the CYP199A2-HaPux complex. The structure is different from the two known Class I CYP-Fdx structures: CYP11A1-Adx and CYP101A1-Pdx. At the protein interface, HaPux residues in the [Fe2S2] cluster-binding loop and the α3 helix, and the C-terminus residue interact with CYP199A2 residues in the proximal loop and the C helix. These residue contacts are consistent with biochemical data on CYP199A2-ferredoxin binding and electron transfer. Electron-tunneling calculations indicate an efficient electron-transfer pathway from the [Fe2S2] cluster to the heme. This new structural model of a CYP-Fdx complex provides the basis for tailoring CYP enzymes for which the cognate ferredoxin is not known, to accept electrons from HaPux and display monooxygenase activity.
dc.relation.ispartofJournal of the American Chemical Societyen
dc.rightsCopyright © 2017 American Chemical Society. This work has been made available online in accordance with the publisher’s policies. This is the author created, accepted version manuscript following peer review and may differ slightly from the final published version. The final published version of this work is available at
dc.subjectQD Chemistryen
dc.titleA structural model of a P450-ferredoxin complex from orientation-selective double electron-electron resonance spectroscopyen
dc.typeJournal articleen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.contributor.institutionUniversity of St Andrews. School of Physics and Astronomyen
dc.description.statusPeer revieweden

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