Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity control

Abstract Cytochrome P450 1A2, as many isoenzymes, can generate multiple metabolites from a single substrate. A loose coupling between substrate binding and oxygen activation makes possible substrate reorientations at the active site prior to catalysis. In the present work, caffeine oxidation to alte...

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Main Authors: Denis Pompon, Luis F. Garcia-Alles, Philippe Urban
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
Subjects:
Online Access:https://doi.org/10.1038/s41598-025-91155-0
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author Denis Pompon
Luis F. Garcia-Alles
Philippe Urban
author_facet Denis Pompon
Luis F. Garcia-Alles
Philippe Urban
author_sort Denis Pompon
collection DOAJ
description Abstract Cytochrome P450 1A2, as many isoenzymes, can generate multiple metabolites from a single substrate. A loose coupling between substrate binding and oxygen activation makes possible substrate reorientations at the active site prior to catalysis. In the present work, caffeine oxidation to alternative bioactive compounds was used to decipher this pluripotency. A model involving two interacting subsites capable of sequentially accommodating one or two caffeine molecules was considered. Molecular dynamics was used to characterize subsite interactions and feed a dedicated geometric encoding of trajectories that was coupled to dimensional reductions and differential machine learning. The two subsites differentially control caffeine orientations and can exchange substrate through a phenylalanine gated mechanism. This exchange can be locked by the presence of a second bound molecule. Complementary roles of subsites in progressively determining the caffeine orientation during its approach to active oxygen were examined. Interestingly, substrate face flipping becomes impaired upon entry into the rather flat active site. This makes the mechanisms that define the orientation of caffeine relative to active oxygen dependent on the substrate face oriented toward heme. Globally, this evidenced that P450 1A2 regioselectivity results from local determinants combined with subsite interactions and caffeine face preselection at a longer distance.
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spelling doaj-art-bedec4216a92447fb49a6ef8a1f5d7152025-08-20T01:57:27ZengNature PortfolioScientific Reports2045-23222025-03-0115111310.1038/s41598-025-91155-0Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity controlDenis Pompon0Luis F. Garcia-Alles1Philippe Urban2Toulouse Biotechnology Institute, Université de Toulouse, CNRS, INRAE, INSAToulouse Biotechnology Institute, Université de Toulouse, CNRS, INRAE, INSAToulouse Biotechnology Institute, Université de Toulouse, CNRS, INRAE, INSAAbstract Cytochrome P450 1A2, as many isoenzymes, can generate multiple metabolites from a single substrate. A loose coupling between substrate binding and oxygen activation makes possible substrate reorientations at the active site prior to catalysis. In the present work, caffeine oxidation to alternative bioactive compounds was used to decipher this pluripotency. A model involving two interacting subsites capable of sequentially accommodating one or two caffeine molecules was considered. Molecular dynamics was used to characterize subsite interactions and feed a dedicated geometric encoding of trajectories that was coupled to dimensional reductions and differential machine learning. The two subsites differentially control caffeine orientations and can exchange substrate through a phenylalanine gated mechanism. This exchange can be locked by the presence of a second bound molecule. Complementary roles of subsites in progressively determining the caffeine orientation during its approach to active oxygen were examined. Interestingly, substrate face flipping becomes impaired upon entry into the rather flat active site. This makes the mechanisms that define the orientation of caffeine relative to active oxygen dependent on the substrate face oriented toward heme. Globally, this evidenced that P450 1A2 regioselectivity results from local determinants combined with subsite interactions and caffeine face preselection at a longer distance.https://doi.org/10.1038/s41598-025-91155-0Deep learningMolecular dynamicGeometric encodingP450CaffeineRegioselectivity
spellingShingle Denis Pompon
Luis F. Garcia-Alles
Philippe Urban
Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity control
Scientific Reports
Deep learning
Molecular dynamic
Geometric encoding
P450
Caffeine
Regioselectivity
title Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity control
title_full Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity control
title_fullStr Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity control
title_full_unstemmed Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity control
title_short Geometry-encoded molecular dynamics enables deep learning insights into P450 regiospecificity control
title_sort geometry encoded molecular dynamics enables deep learning insights into p450 regiospecificity control
topic Deep learning
Molecular dynamic
Geometric encoding
P450
Caffeine
Regioselectivity
url https://doi.org/10.1038/s41598-025-91155-0
work_keys_str_mv AT denispompon geometryencodedmoleculardynamicsenablesdeeplearninginsightsintop450regiospecificitycontrol
AT luisfgarciaalles geometryencodedmoleculardynamicsenablesdeeplearninginsightsintop450regiospecificitycontrol
AT philippeurban geometryencodedmoleculardynamicsenablesdeeplearninginsightsintop450regiospecificitycontrol