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

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

doi:10.1016/j.jinorgbio.2023.112241

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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

, Steven Kelly, Galina I. Lepesheva, John J. Stegeman

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

Swansea University Authors: David Lamb , Steven Kelly

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DOI (Published version): 10.1016/j.jinorgbio.2023.112241

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. The functional significance of these cavities is not known.</abstract><type>Journal Article</type><journal>Journal of Inorganic Biochemistry</journal><volume>245</volume><journalNumber/><paginationStart>112241</paginationStart><paginationEnd/><publisher>Elsevier BV</publisher><placeOfPublication/><isbnPrint/><isbnElectronic/><issnPrint>0162-0134</issnPrint><issnElectronic/><keywords>Adaptation; CYP51; Cytochrome P450; Deep sea; Fish; Pressure.</keywords><publishedDay>31</publishedDay><publishedMonth>8</publishedMonth><publishedYear>2023</publishedYear><publishedDate>2023-08-31</publishedDate><doi>10.1016/j.jinorgbio.2023.112241</doi><url/><notes/><college>COLLEGE NANME</college><department>Medical School</department><CollegeCode>COLLEGE CODE</CollegeCode><DepartmentCode>MEDS</DepartmentCode><institution>Swansea University</institution><apcterm/><funders>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.). 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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 Steven Kelly Steven Kelly true false 2023-06-06 MEDS 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 COLLEGE NANME Medical School COLLEGE CODE MEDS 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.). 2024-07-29T14:15:40.3360139 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 3 Galina I. Lepesheva 4 John J. Stegeman 5 63599__28163__ba766c0239ed4fc5814caca881d35a7e.pdf 63599.AAM.pdf 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_title	Faculty of Medicine, Health and Life Sciences
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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
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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	2023-08-31T14:15:39Z
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Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s

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