Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides

Murarka, Vidhi C; Kim, Jenny S; Lamb, David; Kelly, Steven L; Poulos, Thomas L; Follmer, Alec H

doi:10.1016/j.jbc.2025.108185

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Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides

Vidhi C Murarka, Jenny S Kim, David Lamb

, Steven L Kelly, Thomas L Poulos, Alec H Follmer

The Journal of Biological Chemistry, Volume: 301, Issue: 3, Start page: 108185

Swansea University Author: David Lamb

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DOI (Published version): 10.1016/j.jbc.2025.108185

Abstract

NysL, a cytochrome P450 monooxygenase from the Gram-positive bacterium Streptomyces noursei, catalyzes the C10 hydroxylation of 10-deoxynystain to nystatin A1, a clinically important antifungal. In this study, we present the 2.0 Å resolution crystal structure of NysL bound to nystatin A1. The struct...

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Published in:	The Journal of Biological Chemistry
ISSN:	0021-9258 1083-351X
Published:	Elsevier Inc 2025
Online Access:	Check full text
URI:	https://cronfa.swan.ac.uk/Record/cronfa68750

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spelling	2025-02-18T11:56:56.6262957 v2 68750 2025-01-29 Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides 1dc64e55c2c28d107ef7c3db984cccd2 0000-0001-5446-2997 David Lamb David Lamb true false 2025-01-29 MEDS NysL, a cytochrome P450 monooxygenase from the Gram-positive bacterium Streptomyces noursei, catalyzes the C10 hydroxylation of 10-deoxynystain to nystatin A1, a clinically important antifungal. In this study, we present the 2.0 Å resolution crystal structure of NysL bound to nystatin A1. The structure of this complex provides key insights into the structural elements that dictate the regio- and stereo-selective oxidation of large 20-44-membered macrolide substrates. The closely related AmphL operates on a similar 38-member macrolide but oxidizes C8 rather than C10. This difference requires that the substrate for AmphL penetrate further into the active site relative to NysL. The depth of substrate penetration is controlled by interactions between an area of the substrate binding pocket deemed the “back-wall” and the hemiketal ring of the macrolide substrate. Journal Article The Journal of Biological Chemistry 301 3 108185 Elsevier Inc 0021-9258 1083-351X cytochrome P450; substrate specificity; antifungal; crystal structure 1 3 2025 2025-03-01 10.1016/j.jbc.2025.108185 COLLEGE NANME Medical School COLLEGE CODE MEDS Swansea University Another institution paid the OA fee This work was supported by NIH grant GM131920 (T.L.P.). A.H.F. acknowledges partial support from GM120349 (Andrew S. Borovik). The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH. 2025-02-18T11:56:56.6262957 2025-01-29T10:16:26.5001093 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science Vidhi C Murarka 1 Jenny S Kim 2 David Lamb 0000-0001-5446-2997 3 Steven L Kelly 4 Thomas L Poulos 5 Alec H Follmer 6 68750__33619__db68ef6b47cc47d88bc4a8afa09d9603.pdf 68750.VOR.pdf 2025-02-18T11:49:24.2431964 Output 2851228 application/pdf Version of Record true © 2025 The Authors. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY-NC-ND license. true eng http://creativecommons.org/licenses/by-nc-nd/4.0/
title	Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides
spellingShingle	Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides David Lamb
title_short	Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides
title_full	Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides
title_fullStr	Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides
title_full_unstemmed	Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides
title_sort	Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides
author_id_str_mv	1dc64e55c2c28d107ef7c3db984cccd2
author_id_fullname_str_mv	1dc64e55c2c28d107ef7c3db984cccd2_***_David Lamb
author	David Lamb
author2	Vidhi C Murarka Jenny S Kim David Lamb Steven L Kelly Thomas L Poulos Alec H Follmer
format	Journal article
container_title	The Journal of Biological Chemistry
container_volume	301
container_issue	3
container_start_page	108185
publishDate	2025
institution	Swansea University
issn	0021-9258 1083-351X
doi_str_mv	10.1016/j.jbc.2025.108185
publisher	Elsevier Inc
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	NysL, a cytochrome P450 monooxygenase from the Gram-positive bacterium Streptomyces noursei, catalyzes the C10 hydroxylation of 10-deoxynystain to nystatin A1, a clinically important antifungal. In this study, we present the 2.0 Å resolution crystal structure of NysL bound to nystatin A1. The structure of this complex provides key insights into the structural elements that dictate the regio- and stereo-selective oxidation of large 20-44-membered macrolide substrates. The closely related AmphL operates on a similar 38-member macrolide but oxidizes C8 rather than C10. This difference requires that the substrate for AmphL penetrate further into the active site relative to NysL. The depth of substrate penetration is controlled by interactions between an area of the substrate binding pocket deemed the “back-wall” and the hemiketal ring of the macrolide substrate.
published_date	2025-03-01T08:34:53Z
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Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides

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