Cargando…

New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition

The fourth enzymatic reaction in the de novo pyrimidine biosynthesis, the oxidation of dihydroorotate to orotate, is catalyzed by dihydroorotate dehydrogenase (DHODH). Enzymes belonging to the DHODH Class II are membrane-bound proteins that use ubiquinones as their electron acceptors. We have design...

Descripción completa

Detalles Bibliográficos
Autores principales: Orozco Rodriguez, Juan Manuel, Wacklin-Knecht, Hanna P., Clifton, Luke A., Bogojevic, Oliver, Leung, Anna, Fragneto, Giovanna, Knecht, Wolfgang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8910288/
https://www.ncbi.nlm.nih.gov/pubmed/35269583
http://dx.doi.org/10.3390/ijms23052437
_version_ 1784666431437144064
author Orozco Rodriguez, Juan Manuel
Wacklin-Knecht, Hanna P.
Clifton, Luke A.
Bogojevic, Oliver
Leung, Anna
Fragneto, Giovanna
Knecht, Wolfgang
author_facet Orozco Rodriguez, Juan Manuel
Wacklin-Knecht, Hanna P.
Clifton, Luke A.
Bogojevic, Oliver
Leung, Anna
Fragneto, Giovanna
Knecht, Wolfgang
author_sort Orozco Rodriguez, Juan Manuel
collection PubMed
description The fourth enzymatic reaction in the de novo pyrimidine biosynthesis, the oxidation of dihydroorotate to orotate, is catalyzed by dihydroorotate dehydrogenase (DHODH). Enzymes belonging to the DHODH Class II are membrane-bound proteins that use ubiquinones as their electron acceptors. We have designed this study to understand the interaction of an N-terminally truncated human DHODH (HsΔ29DHODH) and the DHODH from Escherichia coli (EcDHODH) with ubiquinone (Q(10)) in supported lipid membranes using neutron reflectometry (NR). NR has allowed us to determine in situ, under solution conditions, how the enzymes bind to lipid membranes and to unambiguously resolve the location of Q(10). Q(10) is exclusively located at the center of all of the lipid bilayers investigated, and upon binding, both of the DHODHs penetrate into the hydrophobic region of the outer lipid leaflet towards the Q(10). We therefore show that the interaction between the soluble enzymes and the membrane-embedded Q(10) is mediated by enzyme penetration. We can also show that EcDHODH binds more efficiently to the surface of simple bilayers consisting of 1-palmitoyl, 2-oleoyl phosphatidylcholine, and tetraoleoyl cardiolipin than HsΔ29DHODH, but does not penetrate into the lipids to the same degree. Our results also highlight the importance of Q(10), as well as lipid composition, on enzyme binding.
format Online
Article
Text
id pubmed-8910288
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-89102882022-03-11 New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition Orozco Rodriguez, Juan Manuel Wacklin-Knecht, Hanna P. Clifton, Luke A. Bogojevic, Oliver Leung, Anna Fragneto, Giovanna Knecht, Wolfgang Int J Mol Sci Article The fourth enzymatic reaction in the de novo pyrimidine biosynthesis, the oxidation of dihydroorotate to orotate, is catalyzed by dihydroorotate dehydrogenase (DHODH). Enzymes belonging to the DHODH Class II are membrane-bound proteins that use ubiquinones as their electron acceptors. We have designed this study to understand the interaction of an N-terminally truncated human DHODH (HsΔ29DHODH) and the DHODH from Escherichia coli (EcDHODH) with ubiquinone (Q(10)) in supported lipid membranes using neutron reflectometry (NR). NR has allowed us to determine in situ, under solution conditions, how the enzymes bind to lipid membranes and to unambiguously resolve the location of Q(10). Q(10) is exclusively located at the center of all of the lipid bilayers investigated, and upon binding, both of the DHODHs penetrate into the hydrophobic region of the outer lipid leaflet towards the Q(10). We therefore show that the interaction between the soluble enzymes and the membrane-embedded Q(10) is mediated by enzyme penetration. We can also show that EcDHODH binds more efficiently to the surface of simple bilayers consisting of 1-palmitoyl, 2-oleoyl phosphatidylcholine, and tetraoleoyl cardiolipin than HsΔ29DHODH, but does not penetrate into the lipids to the same degree. Our results also highlight the importance of Q(10), as well as lipid composition, on enzyme binding. MDPI 2022-02-23 /pmc/articles/PMC8910288/ /pubmed/35269583 http://dx.doi.org/10.3390/ijms23052437 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Orozco Rodriguez, Juan Manuel
Wacklin-Knecht, Hanna P.
Clifton, Luke A.
Bogojevic, Oliver
Leung, Anna
Fragneto, Giovanna
Knecht, Wolfgang
New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition
title New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition
title_full New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition
title_fullStr New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition
title_full_unstemmed New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition
title_short New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition
title_sort new insights into the interaction of class ii dihydroorotate dehydrogenases with ubiquinone in lipid bilayers as a function of lipid composition
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8910288/
https://www.ncbi.nlm.nih.gov/pubmed/35269583
http://dx.doi.org/10.3390/ijms23052437
work_keys_str_mv AT orozcorodriguezjuanmanuel newinsightsintotheinteractionofclassiidihydroorotatedehydrogenaseswithubiquinoneinlipidbilayersasafunctionoflipidcomposition
AT wacklinknechthannap newinsightsintotheinteractionofclassiidihydroorotatedehydrogenaseswithubiquinoneinlipidbilayersasafunctionoflipidcomposition
AT cliftonlukea newinsightsintotheinteractionofclassiidihydroorotatedehydrogenaseswithubiquinoneinlipidbilayersasafunctionoflipidcomposition
AT bogojevicoliver newinsightsintotheinteractionofclassiidihydroorotatedehydrogenaseswithubiquinoneinlipidbilayersasafunctionoflipidcomposition
AT leunganna newinsightsintotheinteractionofclassiidihydroorotatedehydrogenaseswithubiquinoneinlipidbilayersasafunctionoflipidcomposition
AT fragnetogiovanna newinsightsintotheinteractionofclassiidihydroorotatedehydrogenaseswithubiquinoneinlipidbilayersasafunctionoflipidcomposition
AT knechtwolfgang newinsightsintotheinteractionofclassiidihydroorotatedehydrogenaseswithubiquinoneinlipidbilayersasafunctionoflipidcomposition