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A structural model of a P450-ferredoxin complex from orientation-selective double electron-electron resonance spectroscopy
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dc.contributor.author | Bowen, Alice M. | |
dc.contributor.author | Johnson, Eachan O. D. | |
dc.contributor.author | Mercuri, Francesco | |
dc.contributor.author | Hoskins, Nicola J. | |
dc.contributor.author | Qiao, Ruihong | |
dc.contributor.author | McCullagh, James S. O. | |
dc.contributor.author | Lovett, Janet E. | |
dc.contributor.author | Bell, Stephen G. | |
dc.contributor.author | Zhou, Weihong | |
dc.contributor.author | Timmel, Christiane R. | |
dc.contributor.author | Wong, Luet Lok | |
dc.contributor.author | Harmer, Jeffrey R. | |
dc.date.accessioned | 2018-12-21T00:35:08Z | |
dc.date.available | 2018-12-21T00:35:08Z | |
dc.date.issued | 2018-02-21 | |
dc.identifier | 252001352 | |
dc.identifier | 430d6bcc-8c18-45fd-9f50-cade6f1c013e | |
dc.identifier | 85042365238 | |
dc.identifier | 000426143800024 | |
dc.identifier.citation | Bowen , 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 . https://doi.org/10.1021/jacs.7b11056 | en |
dc.identifier.issn | 0002-7863 | |
dc.identifier.other | ORCID: /0000-0002-3561-450X/work/45525136 | |
dc.identifier.uri | https://hdl.handle.net/10023/16747 | |
dc.description | This 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.abstract | Cytochrome 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.format.extent | 14 | |
dc.format.extent | 2551885 | |
dc.language.iso | eng | |
dc.relation.ispartof | Journal of the American Chemical Society | en |
dc.subject | QD Chemistry | en |
dc.subject | DAS | en |
dc.subject | BDC | en |
dc.subject | R2C | en |
dc.subject.lcc | QD | en |
dc.title | A structural model of a P450-ferredoxin complex from orientation-selective double electron-electron resonance spectroscopy | en |
dc.type | Journal article | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.contributor.institution | University of St Andrews. School of Physics and Astronomy | en |
dc.identifier.doi | https://doi.org/10.1021/jacs.7b11056 | |
dc.description.status | Peer reviewed | en |
dc.date.embargoedUntil | 2018-12-21 |
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