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Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s

Jared V. Goldstone, David Lamb Orcid Logo, Steven Kelly Orcid Logo, Galina I. Lepesheva, John J. Stegeman

Journal of Inorganic Biochemistry, Volume: 245, Start page: 112241

Swansea University Authors: David Lamb Orcid Logo, Steven Kelly Orcid Logo

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Abstract

Cytochromes P450 enzymes involved in the metabolism of endogenous and xenobiotic substrates, provide an excellent model system to study how membrane proteins with unique functions have catalytically adapted through evolution. Molecular adaptation of deep-sea proteins to high hydrostatic pressure rem...

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Published in: Journal of Inorganic Biochemistry
ISSN: 0162-0134
Published: Elsevier BV 2023
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URI: https://cronfa.swan.ac.uk/Record/cronfa63599
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Molecular adaptation of deep-sea proteins to high hydrostatic pressure remains poorly understood. Herein, we have characterized recombinant cytochrome P450 sterol 14-demethylase (CYP51), an essential enzyme of cholesterol biosynthesis, from an abyssal fish species, Coryphaenoides armatus. C. armatus CYP51 was heterologously expressed in Escherichia coli following N-terminal truncation and purified to homogeneity. Recombinant C. armatus CYP51 bound its sterol substrate lanosterol giving a Type I binding spectra (KD 15 μM) and catalyzed lanosterol 14-demethylation turnover at 5.8 nmol/min/nmol P450. C. armatus CYP51 also bound the azole antifungals ketoconazole (KD 0.12 μM ) and propiconazole (KD 0.54 μM) as determined by Type II absorbance spectra. Comparison of C. armatus CYP51 primary sequence and modeled structures with other CYP51s identified amino acid substitutions that may confer an ability to function under pressures of the deep sea and revealed heretofore undescribed internal cavities in human and other non-deep sea CYP51s. 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spelling v2 63599 2023-06-06 Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s 1dc64e55c2c28d107ef7c3db984cccd2 0000-0001-5446-2997 David Lamb David Lamb true false b17cebaf09b4d737b9378a3581e3de93 0000-0001-7991-5040 Steven Kelly Steven Kelly true false 2023-06-06 BMS Cytochromes P450 enzymes involved in the metabolism of endogenous and xenobiotic substrates, provide an excellent model system to study how membrane proteins with unique functions have catalytically adapted through evolution. Molecular adaptation of deep-sea proteins to high hydrostatic pressure remains poorly understood. Herein, we have characterized recombinant cytochrome P450 sterol 14-demethylase (CYP51), an essential enzyme of cholesterol biosynthesis, from an abyssal fish species, Coryphaenoides armatus. C. armatus CYP51 was heterologously expressed in Escherichia coli following N-terminal truncation and purified to homogeneity. Recombinant C. armatus CYP51 bound its sterol substrate lanosterol giving a Type I binding spectra (KD 15 μM) and catalyzed lanosterol 14-demethylation turnover at 5.8 nmol/min/nmol P450. C. armatus CYP51 also bound the azole antifungals ketoconazole (KD 0.12 μM ) and propiconazole (KD 0.54 μM) as determined by Type II absorbance spectra. Comparison of C. armatus CYP51 primary sequence and modeled structures with other CYP51s identified amino acid substitutions that may confer an ability to function under pressures of the deep sea and revealed heretofore undescribed internal cavities in human and other non-deep sea CYP51s. The functional significance of these cavities is not known. Journal Article Journal of Inorganic Biochemistry 245 112241 Elsevier BV 0162-0134 Adaptation; CYP51; Cytochrome P450; Deep sea; Fish; Pressure. 31 8 2023 2023-08-31 10.1016/j.jinorgbio.2023.112241 http://dx.doi.org/10.1016/j.jinorgbio.2023.112241 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University Support for this research was provided by the Woods Hole Center for Oceans and Human Health (the U.S. Oceans and Human Health Program (NIH grant P01ES028938 and National Science Foundation grant OCE-1840381) (J.J.S.), the Ocean Vision 2030 Fund and its generous donors through the Woods Hole Oceanographic Institution (J.V.G.), the Vertebrate Genome Nomenclature Committee NIH U24HG003345 (J.V.G.). Funding at Swansea University was supported by the European Regional Development Fund/ Welsh European Funding Office via the BEACON project (S.L.K.) and a UK-US Fulbright Scholarship (D.C.L.). Funding at Vanderbilt University was supported by NIH grant R01 GM067871 (G.I.L.). 2023-07-24T16:32:44.3279377 2023-06-06T16:45:27.1811126 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science Jared V. Goldstone 1 David Lamb 0000-0001-5446-2997 2 Steven Kelly 0000-0001-7991-5040 3 Galina I. Lepesheva 4 John J. Stegeman 5 Under embargo Under embargo 2023-07-24T16:09:48.9447332 Output 357018 application/pdf Accepted Manuscript true 2024-04-28T00:00:00.0000000 Distributed under the terms of a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND 4.0). true eng https://creativecommons.org/licenses/by-nc-nd/4.0/
title Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s
spellingShingle Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s
David Lamb
Steven Kelly
title_short Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s
title_full Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s
title_fullStr Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s
title_full_unstemmed Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s
title_sort Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s
author_id_str_mv 1dc64e55c2c28d107ef7c3db984cccd2
b17cebaf09b4d737b9378a3581e3de93
author_id_fullname_str_mv 1dc64e55c2c28d107ef7c3db984cccd2_***_David Lamb
b17cebaf09b4d737b9378a3581e3de93_***_Steven Kelly
author David Lamb
Steven Kelly
author2 Jared V. Goldstone
David Lamb
Steven Kelly
Galina I. Lepesheva
John J. Stegeman
format Journal article
container_title Journal of Inorganic Biochemistry
container_volume 245
container_start_page 112241
publishDate 2023
institution Swansea University
issn 0162-0134
doi_str_mv 10.1016/j.jinorgbio.2023.112241
publisher Elsevier BV
college_str Faculty of Medicine, Health and Life Sciences
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hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Biomedical Science{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Biomedical Science
url http://dx.doi.org/10.1016/j.jinorgbio.2023.112241
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description Cytochromes P450 enzymes involved in the metabolism of endogenous and xenobiotic substrates, provide an excellent model system to study how membrane proteins with unique functions have catalytically adapted through evolution. Molecular adaptation of deep-sea proteins to high hydrostatic pressure remains poorly understood. Herein, we have characterized recombinant cytochrome P450 sterol 14-demethylase (CYP51), an essential enzyme of cholesterol biosynthesis, from an abyssal fish species, Coryphaenoides armatus. C. armatus CYP51 was heterologously expressed in Escherichia coli following N-terminal truncation and purified to homogeneity. Recombinant C. armatus CYP51 bound its sterol substrate lanosterol giving a Type I binding spectra (KD 15 μM) and catalyzed lanosterol 14-demethylation turnover at 5.8 nmol/min/nmol P450. C. armatus CYP51 also bound the azole antifungals ketoconazole (KD 0.12 μM ) and propiconazole (KD 0.54 μM) as determined by Type II absorbance spectra. Comparison of C. armatus CYP51 primary sequence and modeled structures with other CYP51s identified amino acid substitutions that may confer an ability to function under pressures of the deep sea and revealed heretofore undescribed internal cavities in human and other non-deep sea CYP51s. The functional significance of these cavities is not known.
published_date 2023-08-31T16:32:12Z
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