{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T17:22:30Z","timestamp":1777051350455,"version":"3.51.4"},"reference-count":36,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T00:00:00Z","timestamp":1671753600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FCT\/MCTES, Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia and Minist\u00e9rio da Ci\u00eancia, Tecnologia e Ensino Superior","award":["UIDB\/50006\/2020"],"award-info":[{"award-number":["UIDB\/50006\/2020"]}]},{"name":"FCT\/MCTES, Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia and Minist\u00e9rio da Ci\u00eancia, Tecnologia e Ensino Superior","award":["UIDP\/50006\/2020"],"award-info":[{"award-number":["UIDP\/50006\/2020"]}]},{"name":"FCT\/MCTES is also acknowledged for supporting the National Portuguese NMR Network","award":["ROTEIRO\/0031\/2013"],"award-info":[{"award-number":["ROTEIRO\/0031\/2013"]}]},{"name":"FCT\/MCTES is also acknowledged for supporting the National Portuguese NMR Network","award":["PINFRA\/22161\/2016"],"award-info":[{"award-number":["PINFRA\/22161\/2016"]}]},{"name":"FCT\/MCTES is also acknowledged for supporting the National Portuguese NMR Network","award":["UIDB\/04326\/2020"],"award-info":[{"award-number":["UIDB\/04326\/2020"]}]},{"name":"FCT\/MCTES is also acknowledged for supporting the National Portuguese NMR Network","award":["PTDC\/QUI-COL\/32351\/2017"],"award-info":[{"award-number":["PTDC\/QUI-COL\/32351\/2017"]}]},{"name":"FCT for funding through the Scientific Employment Stimulus - Individual Call","award":["CEECIND\/ 00466\/2017"],"award-info":[{"award-number":["CEECIND\/ 00466\/2017"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Cytochrome b5 reductase (Cb5R) is a flavoprotein that participates in the reduction of multiple biological redox partners. Co-localization of this protein with nitric oxide sources has been observed in neurons. In addition, the generation of superoxide anion radical by Cb5R has been observed. A search for specific inhibitors of Cb5R to understand the role of this protein in these new functions has been initiated. Previous studies have shown the ability of different flavonoids to inhibit Cb5R. Anthocyanins are a subgroup of flavonoids responsible for most red and blue colors found in flowers and fruits. Although usually represented by the flavylium cation form, these species are only stable at rather acidic pH values (pH \u2264 1). At higher pH values, the flavylium cation is involved in a dynamic reaction network comprising different neutral species with the potential ability to inhibit the activities of Cb5R. This study aims to provide insights into the molecular mechanism of interaction between flavonoids and Cb5R using flavylium salts as dynamic inhibitors. The outcome of this study might lead to the design of improved specific enzyme inhibitors in the future.<\/jats:p>","DOI":"10.3390\/molecules28010123","type":"journal-article","created":{"date-parts":[[2022,12,27]],"date-time":"2022-12-27T02:53:11Z","timestamp":1672109591000},"page":"123","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["The Use of Flavylium Salts as Dynamic Inhibitor Moieties for Human Cb5R"],"prefix":"10.3390","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4893-2205","authenticated-orcid":false,"given":"Oscar H.","family":"Mart\u00ednez-Costa","sequence":"first","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Universidad Aut\u00f3noma de Madrid (UAM), C\/Arturo Duperier 4, 28029-Madrid, Spain"},{"name":"Instituto de Investigaciones Biom\u00e9dicas \u2018Alberto Sols\u2019 (CSIC-UAM), C\/Arturo Duperier 4, 28029-Madrid, Spain"}]},{"given":"Laura","family":"Rodrigues-Miranda","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Universidad Aut\u00f3noma de Madrid (UAM), C\/Arturo Duperier 4, 28029-Madrid, Spain"}]},{"given":"Sofia M.","family":"Clemente","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Associado Para a Qu\u00edmica Verde (LAQV), Rede de Qu\u00edmica e Tecnologia (REQUIMTE), Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1333-9076","authenticated-orcid":false,"given":"Ant\u00f3nio Jorge","family":"Parola","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Associado Para a Qu\u00edmica Verde (LAQV), Rede de Qu\u00edmica e Tecnologia (REQUIMTE), Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0121-3695","authenticated-orcid":false,"given":"Nuno","family":"Basilio","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Associado Para a Qu\u00edmica Verde (LAQV), Rede de Qu\u00edmica e Tecnologia (REQUIMTE), Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9602-4848","authenticated-orcid":false,"given":"Alejandro K.","family":"Samhan-Arias","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Universidad Aut\u00f3noma de Madrid (UAM), C\/Arturo Duperier 4, 28029-Madrid, Spain"},{"name":"Instituto de Investigaciones Biom\u00e9dicas \u2018Alberto Sols\u2019 (CSIC-UAM), C\/Arturo Duperier 4, 28029-Madrid, Spain"},{"name":"Laborat\u00f3rio Associado Para a Qu\u00edmica Verde (LAQV), Rede de Qu\u00edmica e Tecnologia (REQUIMTE), Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.freeradbiomed.2014.04.033","article-title":"Purified NADH-Cytochrome B5 Reductase Is a Novel Superoxide Anion Source Inhibited by Apocynin: Sensitivity to Nitric Oxide and Peroxynitrite","volume":"73","year":"2014","journal-title":"Free Radic. Biol. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/j.abb.2007.10.004","article-title":"Reduction of Ascorbate Free Radical by the Plasma Membrane of Synaptic Terminals from Rat Brain","volume":"469","author":"Duarte","year":"2008","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1016\/j.bbrc.2009.08.076","article-title":"Hydrogen Sulfide Is a Reversible Inhibitor of the NADH Oxidase Activity of Synaptic Plasma Membranes","volume":"388","year":"2009","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1366","DOI":"10.1021\/tx060106t","article-title":"Reductive Detoxification of Arylhydroxylamine Carcinogens by Human NADH Cytochrome B5 Reductase and Cytochrome B5","volume":"19","author":"Kurian","year":"2006","journal-title":"Chem. Res. Toxicol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1124\/mol.112.084616","article-title":"Evidence That Cytochrome B5 and Cytochrome B5 Reductase Can Act as Sole Electron Donors to the Hepatic Cytochrome P450 System","volume":"83","author":"Henderson","year":"2013","journal-title":"Mol. Pharmacol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"148134","DOI":"10.1016\/j.bbabio.2019.148134","article-title":"Human Erythrocytes Exposure to Juglone Leads to an Increase of Superoxide Anion Production Associated with Cytochrome B5 Reductase Uncoupling","volume":"1861","author":"Nogueira","year":"2020","journal-title":"Biochim. Et Biophys. Acta Bioenerg."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1016\/j.bbabio.2017.10.005","article-title":"Topography of Human Cytochrome B5\/Cytochrome B5 Reductase Interacting Domain and Redox Alterations upon Complex Formation","volume":"1859","author":"Almeida","year":"2018","journal-title":"Biochim. Biophys. Acta Bioenerg."},{"key":"ref_8","unstructured":"Thom, R. (2014). Cytochrome B5 as a Pleitropic Metabolic Modular in Mammalian Cells, Nova Publishers. Protein biochemistry, synthesis, structure and cellular functions."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Guti\u00e9rrez-Merino, C., Mart\u00ednez-Costa, O.H., Monsalve, M., and Samhan-Arias, A.K. (2021). Structural Features of Cytochrome B5-Cytochrome B5 Reductase Complex Formation and Implications for the Intramolecular Dynamics of Cytochrome B5 Reductase. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23010118"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.mcn.2008.08.013","article-title":"Clustering of Plasma Membrane-Bound Cytochrome B5 Reductase within \u201clipid Raft\u201d Microdomains of the Neuronal Plasma Membrane","volume":"40","year":"2009","journal-title":"Mol. Cell. Neurosci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.ceca.2014.06.002","article-title":"Caveolin-Rich Lipid Rafts of the Plasma Membrane of Mature Cerebellar Granule Neurons Are Microcompartments for Calcium\/Reactive Oxygen and Nitrogen Species Cross-Talk Signaling","volume":"56","year":"2014","journal-title":"Cell Calcium"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"61","DOI":"10.29245\/2572.942X\/2016\/6.1066","article-title":"Biochemical and Anatomical Basis of Brain Dysfunctions Caused by Cytochrome B5 Reductase Deficiency or Dysregulation","volume":"1","year":"2016","journal-title":"J. Neurol. Neuromedicine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1161\/CIRCRESAHA.117.310705","article-title":"Cytochrome B5 Reductase 3 Modulates Soluble Guanylate Cyclase Redox State and CGMP Signaling","volume":"121","author":"Rahaman","year":"2017","journal-title":"Circ. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"C254","DOI":"10.1152\/ajpcell.00315.2016","article-title":"Nitric Oxide: What\u2019s New to NO?","volume":"312","author":"Ghimire","year":"2017","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.redox.2017.11.021","article-title":"Cytochrome B5 Reductase Is the Component from Neuronal Synaptic Plasma Membrane Vesicles That Generates Superoxide Anion upon Stimulation by Cytochrome c","volume":"15","author":"Fortalezas","year":"2018","journal-title":"Redox Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/S0304-3835(99)00139-1","article-title":"Free Radical Generation from Heterocyclic Amines by Cytochrome B5 Reductase in the Presence of NADH","volume":"143","author":"Maeda","year":"1999","journal-title":"Cancer Lett."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Shalaby, E. (2019). Diseases Related to Types of Free Radicals, IntechOpen. Antioxidants.","DOI":"10.5772\/intechopen.77838"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.2174\/092986711795029735","article-title":"Neuroprotective Actions of Flavonoids","volume":"18","author":"Lagoa","year":"2011","journal-title":"Curr. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e47","DOI":"10.1017\/jns.2016.41","article-title":"Flavonoids: An Overview","volume":"5","author":"Panche","year":"2016","journal-title":"J. Nutr. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1039\/C1CS15126F","article-title":"Chemistry and Applications of Flavylium Compounds: A Handful of Colours","volume":"41","author":"Pina","year":"2012","journal-title":"Chem. Soc. Rev."},{"key":"ref_21","first-page":"33","article-title":"Molecular Construction of NADH-Cytochrome B5 Reductase Inhibition by Flavonoids and Chemical Basis of Difference in Inhibition Potential: Molecular Dynamics Simulation Study","volume":"2","author":"Verma","year":"2012","journal-title":"J. App. Pharm. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.cbi.2012.04.003","article-title":"Inhibitory Effects of Dietary Flavonoids on Purified Hepatic NADH-Cytochrome B5 Reductase: Structure-Activity Relationships","volume":"197","year":"2012","journal-title":"Chem. Biol. Interact."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1416","DOI":"10.1021\/acs.chemrev.1c00399","article-title":"Natural and Synthetic Flavylium-Based Dyes: The Chemistry Behind the Color","volume":"122","author":"Cruz","year":"2022","journal-title":"Chem. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1039\/b9pp00035f","article-title":"Characterization of the 4\u2032-Hydroxynaphthoflavylium Network of Chemical Reactions","volume":"9","author":"Gavara","year":"2010","journal-title":"Photochem. Photobiol. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.1002\/cplu.201500304","article-title":"Host-Guest Complexes of Flavylium Cations and Cucurbit [7]Uril: The Influence of Flavylium Substituents on the Structure and Stability of the Complex","volume":"80","author":"Petrov","year":"2015","journal-title":"ChemPlusChem"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1039\/c3pp25347c","article-title":"Synthetic Analogues of Anthocyanins as Sensitizers for Dye-Sensitized Solar Cells","volume":"12","author":"Calogero","year":"2013","journal-title":"Photochem. Photobiol. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: Improving the Speed and Accuracy of Docking with a New Scoring Function, Efficient Optimization and Multithreading","volume":"31","author":"Trott","year":"2010","journal-title":"J. Comput. Chem."},{"key":"ref_28","first-page":"57","article-title":"Python: A Programming Language for Software Integration and Development","volume":"17","author":"Sanner","year":"1999","journal-title":"J. Mol. Graph. Model"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2934","DOI":"10.1016\/j.jprot.2011.12.007","article-title":"Stimulation and Clustering of Cytochrome B5 Reductase in Caveolin-Rich Lipid Microdomains Is an Early Event in Oxidative Stress-Mediated Apoptosis of Cerebellar Granule Neurons","volume":"75","author":"Yanamala","year":"2012","journal-title":"J. Proteom."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1111\/j.1471-4159.2009.06331.x","article-title":"Kaempferol Protects against Rat Striatal Degeneration Induced by 3-Nitropropionic Acid","volume":"111","author":"Lagoa","year":"2009","journal-title":"J. Neurochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.freeradbiomed.2004.04.002","article-title":"Kaempferol Blocks Oxidative Stress in Cerebellar Granule Cells and Reveals a Key Role for Reactive Oxygen Species Production at the Plasma Membrane in the Commitment to Apoptosis","volume":"37","year":"2004","journal-title":"Free Radic. Biol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.brainres.2007.08.087","article-title":"Blood Micromolar Concentrations of Kaempferol Afford Protection against Ischemia\/Reperfusion-Induced Damage in Rat Brain","volume":"1182","author":"Sun","year":"2007","journal-title":"Brain Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4294","DOI":"10.1016\/j.tet.2016.05.076","article-title":"A Simple Synthesis of 3-Deoxyanthocyanidins and Their O-Glucosides","volume":"72","author":"Mora","year":"2016","journal-title":"Tetrahedron"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"13574","DOI":"10.1021\/bi0106336","article-title":"The Structure and Biochemistry of NADH-Dependent Cytochrome B5 Reductase Are Now Consistent","volume":"40","author":"Bewley","year":"2001","journal-title":"Biochemistry"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1002\/jcc.20084","article-title":"UCSF Chimera\u2014A Visualization System for Exploratory Research and Analysis","volume":"25","author":"Pettersen","year":"2004","journal-title":"J. Comput. Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/1\/123\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:49:23Z","timestamp":1760147363000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/1\/123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,23]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["molecules28010123"],"URL":"https:\/\/doi.org\/10.3390\/molecules28010123","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,23]]}}}