Cargando…

Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes

This study aimed at searching for the enzymes that are responsible for the higher hydroxylation of flavonols serving as UV-honey guides for pollinating insects on the petals of Asteraceae flowers. To achieve this aim, an affinity-based chemical proteomic approach was developed by relying on the use...

Descripción completa

Detalles Bibliográficos
Autores principales: Kempf, Karl, Kempf, Oxana, Capello, Yoan, Molitor, Christian, Lescoat, Claire, Melhem, Rana, Chaignepain, Stéphane, Génot, Elisabeth, Groppi, Alexis, Nikolski, Macha, Halbwirth, Heidi, Deffieux, Denis, Quideau, Stéphane
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10253404/
https://www.ncbi.nlm.nih.gov/pubmed/37298676
http://dx.doi.org/10.3390/ijms24119724
_version_ 1785056397813088256
author Kempf, Karl
Kempf, Oxana
Capello, Yoan
Molitor, Christian
Lescoat, Claire
Melhem, Rana
Chaignepain, Stéphane
Génot, Elisabeth
Groppi, Alexis
Nikolski, Macha
Halbwirth, Heidi
Deffieux, Denis
Quideau, Stéphane
author_facet Kempf, Karl
Kempf, Oxana
Capello, Yoan
Molitor, Christian
Lescoat, Claire
Melhem, Rana
Chaignepain, Stéphane
Génot, Elisabeth
Groppi, Alexis
Nikolski, Macha
Halbwirth, Heidi
Deffieux, Denis
Quideau, Stéphane
author_sort Kempf, Karl
collection PubMed
description This study aimed at searching for the enzymes that are responsible for the higher hydroxylation of flavonols serving as UV-honey guides for pollinating insects on the petals of Asteraceae flowers. To achieve this aim, an affinity-based chemical proteomic approach was developed by relying on the use of quercetin-bearing biotinylated probes, which were thus designed and synthesized to selectively and covalently capture relevant flavonoid enzymes. Proteomic and bioinformatic analyses of proteins captured from petal microsomes of two Asteraceae species (Rudbeckia hirta and Tagetes erecta) revealed the presence of two flavonol 6-hydroxylases and several additional not fully characterized proteins as candidates for the identification of novel flavonol 8-hydroxylases, as well as relevant flavonol methyl- and glycosyltransferases. Generally speaking, this substrate-based proteome profiling methodology constitutes a powerful tool for the search for unknown (flavonoid) enzymes in plant protein extracts.
format Online
Article
Text
id pubmed-10253404
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-102534042023-06-10 Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes Kempf, Karl Kempf, Oxana Capello, Yoan Molitor, Christian Lescoat, Claire Melhem, Rana Chaignepain, Stéphane Génot, Elisabeth Groppi, Alexis Nikolski, Macha Halbwirth, Heidi Deffieux, Denis Quideau, Stéphane Int J Mol Sci Article This study aimed at searching for the enzymes that are responsible for the higher hydroxylation of flavonols serving as UV-honey guides for pollinating insects on the petals of Asteraceae flowers. To achieve this aim, an affinity-based chemical proteomic approach was developed by relying on the use of quercetin-bearing biotinylated probes, which were thus designed and synthesized to selectively and covalently capture relevant flavonoid enzymes. Proteomic and bioinformatic analyses of proteins captured from petal microsomes of two Asteraceae species (Rudbeckia hirta and Tagetes erecta) revealed the presence of two flavonol 6-hydroxylases and several additional not fully characterized proteins as candidates for the identification of novel flavonol 8-hydroxylases, as well as relevant flavonol methyl- and glycosyltransferases. Generally speaking, this substrate-based proteome profiling methodology constitutes a powerful tool for the search for unknown (flavonoid) enzymes in plant protein extracts. MDPI 2023-06-03 /pmc/articles/PMC10253404/ /pubmed/37298676 http://dx.doi.org/10.3390/ijms24119724 Text en © 2023 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
Kempf, Karl
Kempf, Oxana
Capello, Yoan
Molitor, Christian
Lescoat, Claire
Melhem, Rana
Chaignepain, Stéphane
Génot, Elisabeth
Groppi, Alexis
Nikolski, Macha
Halbwirth, Heidi
Deffieux, Denis
Quideau, Stéphane
Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes
title Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes
title_full Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes
title_fullStr Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes
title_full_unstemmed Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes
title_short Synthesis of Flavonol-Bearing Probes for Chemoproteomic and Bioinformatic Analyses of Asteraceae Petals in Search of Novel Flavonoid Enzymes
title_sort synthesis of flavonol-bearing probes for chemoproteomic and bioinformatic analyses of asteraceae petals in search of novel flavonoid enzymes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10253404/
https://www.ncbi.nlm.nih.gov/pubmed/37298676
http://dx.doi.org/10.3390/ijms24119724
work_keys_str_mv AT kempfkarl synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT kempfoxana synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT capelloyoan synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT molitorchristian synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT lescoatclaire synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT melhemrana synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT chaignepainstephane synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT genotelisabeth synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT groppialexis synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT nikolskimacha synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT halbwirthheidi synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT deffieuxdenis synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes
AT quideaustephane synthesisofflavonolbearingprobesforchemoproteomicandbioinformaticanalysesofasteraceaepetalsinsearchofnovelflavonoidenzymes