Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes

14 pags., 7 figs., 2 tabs. -- Open Access funded by Creative Commons Atribution Licence 4.0

Detalles Bibliográficos
Autores: Arilla-Luna, Sonia, Serrano, Ana, Medina, Milagros
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/206553
Acceso en línea:http://hdl.handle.net/10261/206553
Access Level:acceso abierto
Palabra clave:Prokaryotic FAD synthase
Species-specific traits
Adenylyltransferase activity
Head-to-tail organization
Isothermal titration calorimetry
Site-directed mutagenesis
Biosynthesis
Steady-state kinetics
id ES_3effe2014ecfbdc5bb9d53b1bba9f217
oai_identifier_str oai:digital.csic.es:10261/206553
network_acronym_str ES
network_name_str España
repository_id_str
spelling Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenesArilla-Luna, SoniaSerrano, AnaMedina, MilagrosProkaryotic FAD synthaseSpecies-specific traitsAdenylyltransferase activityHead-to-tail organizationIsothermal titration calorimetrySite-directed mutagenesisBiosynthesisSteady-state kinetics14 pags., 7 figs., 2 tabs. -- Open Access funded by Creative Commons Atribution Licence 4.0Bifunctional FAD synthases (FADSs) catalyze FMN (flavin mononucleotide) and FAD (flavinadenine dinucleotide) biosynthesis at their C-riboflavin kinase (RFK) and N-FMN: adenylyltransferase (FMNAT) modules, respectively. Biophysical properties and requirements for their FMNAT activity differ among species. Here, we evaluate the relevance of the integrity of the binding site of the isoalloxazine of flavinic substrates for FMNAT catalysis in Corynebacterium ammoniagenes FADS (CaFADS). We have substituted P56 and P58, belonging to a conserved motif, as well as L98. These residues shape the isoalloxazine FMNAT site, although they are not expected to directly contact it. All substitutions override enzyme ability to transform substrates at the FMNAT site, although most variants are able to bind them. Spectroscopic properties and thermodynamic parameters for the binding of ligands indicate that mutations alter their interaction modes. Substitutions also modulate binding and kinetic properties at the RFK site, evidencing the crosstalk of different protomers within CaFADS assemblies during catalysis. In conclusion, despite the FMNAT site for the binding of substrates in CaFADS appearing as a wide open cavity, it is finely tuned to provide the competent binding conformation of substrates. In particular, P56, P58 and L98 shape the isoalloxazine site to place the FMN-and FAD-reacting phosphates in optimal geometry for catalysis.This research was founded by 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_17R to M.M.)).Molecular Diversity Preservation InternationalMinisterio de Economía y Competitividad (España)Gobierno de AragónSCOAPConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2020202020192020info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/206553reponame: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-Phttp://dx.doi.org/10.3390/ijms20205083Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2065532026-05-22T06:33:51Z
dc.title.none.fl_str_mv Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
title Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
spellingShingle Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
Arilla-Luna, Sonia
Prokaryotic FAD synthase
Species-specific traits
Adenylyltransferase activity
Head-to-tail organization
Isothermal titration calorimetry
Site-directed mutagenesis
Biosynthesis
Steady-state kinetics
title_short Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
title_full Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
title_fullStr Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
title_full_unstemmed Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
title_sort Specific features for the competent binding of substrates at the FMN adenylyltransferase site of FAD synthase from corynebacterium ammoniagenes
dc.creator.none.fl_str_mv Arilla-Luna, Sonia
Serrano, Ana
Medina, Milagros
author Arilla-Luna, Sonia
author_facet Arilla-Luna, Sonia
Serrano, Ana
Medina, Milagros
author_role author
author2 Serrano, Ana
Medina, Milagros
author2_role author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
Gobierno de Aragón
SCOAP
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Prokaryotic FAD synthase
Species-specific traits
Adenylyltransferase activity
Head-to-tail organization
Isothermal titration calorimetry
Site-directed mutagenesis
Biosynthesis
Steady-state kinetics
topic Prokaryotic FAD synthase
Species-specific traits
Adenylyltransferase activity
Head-to-tail organization
Isothermal titration calorimetry
Site-directed mutagenesis
Biosynthesis
Steady-state kinetics
description 14 pags., 7 figs., 2 tabs. -- Open Access funded by Creative Commons Atribution Licence 4.0
publishDate 2019
dc.date.none.fl_str_mv 2019
2020
2020
2020
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/10261/206553
url http://hdl.handle.net/10261/206553
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
http://dx.doi.org/10.3390/ijms20205083
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Molecular Diversity Preservation International
publisher.none.fl_str_mv Molecular Diversity Preservation International
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
_version_ 1869406604012552192
score 15.81155

Similares en LA Referencia