{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T20:25:35Z","timestamp":1776716735685,"version":"3.51.2"},"reference-count":66,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2023,7,6]],"date-time":"2023-07-06T00:00:00Z","timestamp":1688601600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Portuguese Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia (FCT)","doi-asserted-by":"publisher","award":["UID\/BIM\/0009\/2020"],"award-info":[{"award-number":["UID\/BIM\/0009\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Portuguese Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia (FCT)","doi-asserted-by":"publisher","award":["HLTH-2022-STAYHLTH-02"],"award-info":[{"award-number":["HLTH-2022-STAYHLTH-02"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Portuguese Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia (FCT)","doi-asserted-by":"publisher","award":["101095679"],"award-info":[{"award-number":["101095679"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"name":"European Horizon\u00b4s Research and Innovation Program","award":["UID\/BIM\/0009\/2020"],"award-info":[{"award-number":["UID\/BIM\/0009\/2020"]}]},{"name":"European Horizon\u00b4s Research and Innovation Program","award":["HLTH-2022-STAYHLTH-02"],"award-info":[{"award-number":["HLTH-2022-STAYHLTH-02"]}]},{"name":"European Horizon\u00b4s Research and Innovation Program","award":["101095679"],"award-info":[{"award-number":["101095679"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>A unique cytochrome P450 (CYP) oxidoreductase (CPR) sustains activities of human microsomal CYPs. Its function requires toggling between a closed conformation enabling electron transfers from NADPH to FAD and then FMN cofactors and open conformations forming complexes and transferring electrons to CYPs. We previously demonstrated that distinct features of the hinge region linking the FAD and FMN domain (FD) modulate conformer poses and their interactions with CYPs. Specific FD residues contribute in a CYP isoform-dependent manner to the recognition and electron transfer mechanisms that are additionally modulated by the structure of CYP-bound substrate. To obtain insights into the underlying mechanisms, we analyzed how hinge region and FD mutations influence CYP1A2-mediated caffeine metabolism. Activities, metabolite profiles, regiospecificity and coupling efficiencies were evaluated in regard to the structural features and molecular dynamics of complexes bearing alternate substrate poses at the CYP active site. Studies reveal that FD variants not only modulate CYP activities but surprisingly the regiospecificity of reactions. Computational approaches evidenced that the considered mutations are generally in close contact with residues at the FD\u2013CYP interface, exhibiting induced fits during complexation and modified dynamics depending on caffeine presence and orientation. It was concluded that dynamic coupling between FD mutations, the complex interface and CYP active site exist consistently with the observed regiospecific alterations.<\/jats:p>","DOI":"10.3390\/biom13071083","type":"journal-article","created":{"date-parts":[[2023,7,7]],"date-time":"2023-07-07T01:41:36Z","timestamp":1688694096000},"page":"1083","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Single Mutations in Cytochrome P450 Oxidoreductase Can Alter the Specificity of Human Cytochrome P450 1A2-Mediated Caffeine Metabolism"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3046-1313","authenticated-orcid":false,"given":"Francisco","family":"Esteves","sequence":"first","affiliation":[{"name":"ToxOmics, NOVA Medical School, Faculdade de Ci\u00eancias M\u00e9dicas, NMS|FCM, Universidade NOVA de Lisboa, Campo M\u00e1rtires da P\u00e1tria 130, 1169-056 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6872-9127","authenticated-orcid":false,"given":"Cristina M. M.","family":"Almeida","sequence":"additional","affiliation":[{"name":"iMed.UL (Institute for Medicines and Pharmaceutical Sciences, Portugal), Faculty of Pharmacy, University of Lisboa, Av. Prof. Gama Pinto, 2, 1649-003 Lisbon, Portugal"},{"name":"Laboratory of Bromatology and Water Quality, Faculty of Pharmacy, University of Lisbon, Av. Prof. Gama Pinto, 2, 1649-003 Lisbon, Portugal"}]},{"given":"Sofia","family":"Silva","sequence":"additional","affiliation":[{"name":"Laboratory of Bromatology and Water Quality, Faculty of Pharmacy, University of Lisbon, Av. Prof. Gama Pinto, 2, 1649-003 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5842-6715","authenticated-orcid":false,"given":"In\u00eas","family":"Saldanha","sequence":"additional","affiliation":[{"name":"ToxOmics, NOVA Medical School, Faculdade de Ci\u00eancias M\u00e9dicas, NMS|FCM, Universidade NOVA de Lisboa, Campo M\u00e1rtires da P\u00e1tria 130, 1169-056 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0557-839X","authenticated-orcid":false,"given":"Philippe","family":"Urban","sequence":"additional","affiliation":[{"name":"TBI, Universit\u00e9 de Toulouse, CNRS, INRAE, INSA, 135 Avenue de Rangueil, 31077 Toulouse, CEDEX 04, France"}]},{"given":"Jos\u00e9","family":"Rueff","sequence":"additional","affiliation":[{"name":"ToxOmics, NOVA Medical School, Faculdade de Ci\u00eancias M\u00e9dicas, NMS|FCM, Universidade NOVA de Lisboa, Campo M\u00e1rtires da P\u00e1tria 130, 1169-056 Lisbon, Portugal"}]},{"given":"Denis","family":"Pompon","sequence":"additional","affiliation":[{"name":"TBI, Universit\u00e9 de Toulouse, CNRS, INRAE, INSA, 135 Avenue de Rangueil, 31077 Toulouse, CEDEX 04, France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3560-8302","authenticated-orcid":false,"given":"Gilles","family":"Truan","sequence":"additional","affiliation":[{"name":"TBI, Universit\u00e9 de Toulouse, CNRS, INRAE, INSA, 135 Avenue de Rangueil, 31077 Toulouse, CEDEX 04, France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1362-0076","authenticated-orcid":false,"given":"Michel","family":"Kranendonk","sequence":"additional","affiliation":[{"name":"ToxOmics, NOVA Medical School, Faculdade de Ci\u00eancias M\u00e9dicas, NMS|FCM, Universidade NOVA de Lisboa, Campo M\u00e1rtires da P\u00e1tria 130, 1169-056 Lisbon, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ortiz de Montellano, P.R. 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