Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis

16 p.-9 fig.-3 tab.

Detalles Bibliográficos
Autores: Serrano, Ana, Arilla-Luna, Sonia, Medina, Milagros
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/233124
Acceso en línea:http://hdl.handle.net/10261/233124
Access Level:acceso abierto
Palabra clave:Flavin biosynthesis
Prokaryotic FAD synthase
Eukaryotic ATP:FMN:adenylyltransferase
Aromatic residues
Isoalloxazine
Binding
Rational mutagenesis
Isothermal titration calorimetry
Steady-state kinetics
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spelling Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysisSerrano, AnaArilla-Luna, SoniaMedina, MilagrosFlavin biosynthesisProkaryotic FAD synthaseEukaryotic ATP:FMN:adenylyltransferaseAromatic residuesIsoalloxazineBindingRational mutagenesisIsothermal titration calorimetrySteady-state kinetics16 p.-9 fig.-3 tab.The last step in the biosynthesis of flavin adenine dinucleotide (FAD) is considered a target for the design of antimicrobial drugs because it is carried out by two non-homologous proteins in eukaryotic and prokaryotic organisms. Monofunctional FMN: adenylyltransferases (FMNAT) in Eukarya and FMNAT modules of bifunctional FAD synthases (FADS) in Prokarya belong to different structural families with dissimilar chemistry and binding modes for the substrates. In this study, we analyzed the relevance of the hydrophobic environment of the flavin isoalloxazine in the FMNAT active site of Corynebacterium ammoniagenes FADS (CaFADS) through the mutational analysis of its F62, Y106, and F128 residues. They form the isoalloxazine binding cavity and are highly conserved in the prokaryotic FADS family. The spectroscopic, steady-state kinetics and thermodynamic data presented indicate that distortion of aromaticity at the FMNAT isoalloxazine binding cavity prevents FMN and FAD from correct accommodation in their binding cavity and, as a consequence, decreases the efficiency of the FMNAT activity. Therefore, the side-chains of F62, Y106 and F128 are relevant in the formation of the catalytic competent complex during FMNAT catalysis in CaFADS. The introduced mutations also modulate the activity occurring at the riboflavin kinase (RFK) module of CaFADS, further evidencing the formation of quaternary assemblies during catalysis.The Spanish Agencia Estatal de Investigación and Fondo Europeo de Desarrollo Regional (BIO2016-75183-P AEI/FEDER, UE to M.M.) and the Government of Aragón-FEDER (grupo de Referencia Biología Estructural (E35_20R to M.M.)) funded this research.Peer reviewedMultidisciplinary Digital Publishing InstituteAgencia Estatal de Investigación (España)European CommissionGobierno de AragónSerrano, Ana [0000-0002-7057-0418]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202120212020info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/233124reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-75183-Phttps://doi.org/10.3390/ijms21103738Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2331242026-05-22T06:33:51Z
dc.title.none.fl_str_mv Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
title Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
spellingShingle Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
Serrano, Ana
Flavin biosynthesis
Prokaryotic FAD synthase
Eukaryotic ATP:FMN:adenylyltransferase
Aromatic residues
Isoalloxazine
Binding
Rational mutagenesis
Isothermal titration calorimetry
Steady-state kinetics
title_short Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
title_full Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
title_fullStr Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
title_full_unstemmed Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
title_sort Insights into the FMNAT active site of FADSynthase: aromaticity is essential for flavin bindingand catalysis
dc.creator.none.fl_str_mv Serrano, Ana
Arilla-Luna, Sonia
Medina, Milagros
author Serrano, Ana
author_facet Serrano, Ana
Arilla-Luna, Sonia
Medina, Milagros
author_role author
author2 Arilla-Luna, Sonia
Medina, Milagros
author2_role author
author
dc.contributor.none.fl_str_mv Agencia Estatal de Investigación (España)
European Commission
Gobierno de Aragón
Serrano, Ana [0000-0002-7057-0418]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Flavin biosynthesis
Prokaryotic FAD synthase
Eukaryotic ATP:FMN:adenylyltransferase
Aromatic residues
Isoalloxazine
Binding
Rational mutagenesis
Isothermal titration calorimetry
Steady-state kinetics
topic Flavin biosynthesis
Prokaryotic FAD synthase
Eukaryotic ATP:FMN:adenylyltransferase
Aromatic residues
Isoalloxazine
Binding
Rational mutagenesis
Isothermal titration calorimetry
Steady-state kinetics
description 16 p.-9 fig.-3 tab.
publishDate 2020
dc.date.none.fl_str_mv 2020
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/233124
url http://hdl.handle.net/10261/233124
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-75183-P
https://doi.org/10.3390/ijms21103738
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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