Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis

Abstract Cytochrome P450 reductases (CPRs) are diflavin oxidoreductases that supply electrons to type II cytochrome P450 monooxygenases (CYPs). In addition, it can also reduce other proteins and molecules, including cytochrome c, ferricyanide, and different drugs. Although various CPRs have been fun...

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Autores principales:	Ana C. Ebrecht, Naadia van der Bergh, Susan T. L. Harrison, Martha S. Smit, B. Trevor Sewell, Diederik J. Opperman
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/802f6970e4aa4dd5ac1d70d7584fd111
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spelling	oai:doaj.org-article:802f6970e4aa4dd5ac1d70d7584fd1112021-12-02T13:34:54ZBiochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis10.1038/s41598-019-56516-62045-2322https://doaj.org/article/802f6970e4aa4dd5ac1d70d7584fd1112019-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-56516-6https://doaj.org/toc/2045-2322Abstract Cytochrome P450 reductases (CPRs) are diflavin oxidoreductases that supply electrons to type II cytochrome P450 monooxygenases (CYPs). In addition, it can also reduce other proteins and molecules, including cytochrome c, ferricyanide, and different drugs. Although various CPRs have been functionally and structurally characterized, the overall mechanism and its interaction with different redox acceptors remain elusive. One of the main problems regarding electron transfer between CPRs and CYPs is the so-called “uncoupling”, whereby NAD(P)H derived electrons are lost due to the reduced intermediates’ (FAD and FMN of CPR) interaction with molecular oxygen. Additionally, the decay of the iron-oxygen complex of the CYP can also contribute to loss of reducing equivalents during an unproductive reaction cycle. This phenomenon generates reactive oxygen species (ROS), leading to an inefficient reaction. Here, we present the study of the CPR from Candida tropicalis (CtCPR) lacking the hydrophobic N-terminal part (Δ2–22). The enzyme supports the reduction of cytochrome c and ferricyanide, with an estimated 30% uncoupling during the reactions with cytochrome c. The ROS produced was not influenced by different physicochemical conditions (ionic strength, pH, temperature). The X-ray structures of the enzyme were solved with and without its cofactor, NADPH. Both CtCPR structures exhibited the closed conformation. Comparison with the different solved structures revealed an intricate ionic network responsible for the regulation of the open/closed movement of CtCPR.Ana C. EbrechtNaadia van der BerghSusan T. L. HarrisonMartha S. SmitB. Trevor SewellDiederik J. OppermanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-11 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ana C. Ebrecht Naadia van der Bergh Susan T. L. Harrison Martha S. Smit B. Trevor Sewell Diederik J. Opperman Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis
description	Abstract Cytochrome P450 reductases (CPRs) are diflavin oxidoreductases that supply electrons to type II cytochrome P450 monooxygenases (CYPs). In addition, it can also reduce other proteins and molecules, including cytochrome c, ferricyanide, and different drugs. Although various CPRs have been functionally and structurally characterized, the overall mechanism and its interaction with different redox acceptors remain elusive. One of the main problems regarding electron transfer between CPRs and CYPs is the so-called “uncoupling”, whereby NAD(P)H derived electrons are lost due to the reduced intermediates’ (FAD and FMN of CPR) interaction with molecular oxygen. Additionally, the decay of the iron-oxygen complex of the CYP can also contribute to loss of reducing equivalents during an unproductive reaction cycle. This phenomenon generates reactive oxygen species (ROS), leading to an inefficient reaction. Here, we present the study of the CPR from Candida tropicalis (CtCPR) lacking the hydrophobic N-terminal part (Δ2–22). The enzyme supports the reduction of cytochrome c and ferricyanide, with an estimated 30% uncoupling during the reactions with cytochrome c. The ROS produced was not influenced by different physicochemical conditions (ionic strength, pH, temperature). The X-ray structures of the enzyme were solved with and without its cofactor, NADPH. Both CtCPR structures exhibited the closed conformation. Comparison with the different solved structures revealed an intricate ionic network responsible for the regulation of the open/closed movement of CtCPR.
format	article
author	Ana C. Ebrecht Naadia van der Bergh Susan T. L. Harrison Martha S. Smit B. Trevor Sewell Diederik J. Opperman
author_facet	Ana C. Ebrecht Naadia van der Bergh Susan T. L. Harrison Martha S. Smit B. Trevor Sewell Diederik J. Opperman
author_sort	Ana C. Ebrecht
title	Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis
title_short	Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis
title_full	Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis
title_fullStr	Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis
title_full_unstemmed	Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis
title_sort	biochemical and structural insights into the cytochrome p450 reductase from candida tropicalis
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/802f6970e4aa4dd5ac1d70d7584fd111
work_keys_str_mv	AT anacebrecht biochemicalandstructuralinsightsintothecytochromep450reductasefromcandidatropicalis AT naadiavanderbergh biochemicalandstructuralinsightsintothecytochromep450reductasefromcandidatropicalis AT susantlharrison biochemicalandstructuralinsightsintothecytochromep450reductasefromcandidatropicalis AT marthassmit biochemicalandstructuralinsightsintothecytochromep450reductasefromcandidatropicalis AT btrevorsewell biochemicalandstructuralinsightsintothecytochromep450reductasefromcandidatropicalis AT diederikjopperman biochemicalandstructuralinsightsintothecytochromep450reductasefromcandidatropicalis
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Biochemical and structural insights into the cytochrome P450 reductase from Candida tropicalis

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