Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners

18 pags., 4 figs., 5 tabs. -- This article belongs to the Special Issue Folding and Design of α-Helical Proteins and Peptides: Theory Meets Nanomaterials, Biotechnology and Health

Detalhes bibliográficos
Autores: Neira, José L., Ortega-Alarcón, David, Rizzuti, Bruno, Palomino-Schätzlein, M., Velázquez-Campoy, Adrián, Falcó, Alberto
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/256259
Acesso em linha:http://hdl.handle.net/10261/256259
Access Level:acceso abierto
Palavra-chave:Binding
Circular dichroism
Peptides
Isothermal titration calorimetry
NMR
Fluorescence
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spelling Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partnersNeira, José L.Ortega-Alarcón, DavidRizzuti, BrunoPalomino-Schätzlein, M.Velázquez-Campoy, AdriánFalcó, AlbertoBindingCircular dichroismPeptidesIsothermal titration calorimetryNMRFluorescence18 pags., 4 figs., 5 tabs. -- This article belongs to the Special Issue Folding and Design of α-Helical Proteins and Peptides: Theory Meets Nanomaterials, Biotechnology and HealthThe phosphoenolpyruvate-dependent phosphotransferase system (PTS) modulates the preferential use of sugars in bacteria. The first proteins in the cascade are common to all organisms (EI and HPr). The active site of HPr involves a histidine (His15) located immediately before the beginning of the first α-helix. The regulator of sigma D (Rsd) protein also binds to HPr. The region of HPr comprising residues Gly9-Ala30 (HPr), involving the first α-helix (Ala16-Thr27) and the preceding active site loop, binds to both the N-terminal region of EI and intact Rsd. HPr is mainly disordered. We attempted to improve the affinity of HPr to both proteins by mutating its sequence to increase its helicity. We designed peptides that led to a marginally larger population in solution of the helical structure of HPr. Molecular simulations also suggested a modest increment in the helical population of mutants, when compared to the wild-type. The mutants, however, were bound with a less favorable affinity than the wild-type to both the N-terminal of EI (EIN) or Rsd, as tested by isothermal titration calorimetry and fluorescence. Furthermore, mutants showed lower antibacterial properties against Staphylococcus aureus than the wild-type peptide. The refore, we concluded that in HPr, a compromise between binding to its partners and residual structure at the active site must exist to carry out its function.This research was funded by the Spanish Ministry of Economy and Competitiveness and European ERDF Funds (MCIU/AEI/FEDER, EU) (RTI2018-097991-B-I00 to J.L.N., BFU2016-78232-P to A.V.-C., BES-2017-080739 to D.O.-A., and RTI2018-101969-J-I00 to A.F.). The NMR equipment used in this work was funded by Generalitat Valenciana (Spain) and cofinanced with ERDF funds (OP ERDF of Comunitat Valenciana (Spain) 2014–2020).Molecular Diversity Preservation InternationalMinisterio de Economía y Competitividad (España)Generalitat ValencianaConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2021202120212021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/256259reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097991-B-I00info:eu-repo/grantAgreement/MINECO//BFU2016-78232-Phttp://dx.doi.org/10.3390/ijms221910805Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2562592026-05-22T06:33:51Z
dc.title.none.fl_str_mv Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
title Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
spellingShingle Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
Neira, José L.
Binding
Circular dichroism
Peptides
Isothermal titration calorimetry
NMR
Fluorescence
title_short Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
title_full Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
title_fullStr Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
title_full_unstemmed Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
title_sort Residual helicity at the active site of the histidine phosphocarrier, HPr, modulates binding affinity to its natural partners
dc.creator.none.fl_str_mv Neira, José L.
Ortega-Alarcón, David
Rizzuti, Bruno
Palomino-Schätzlein, M.
Velázquez-Campoy, Adrián
Falcó, Alberto
author Neira, José L.
author_facet Neira, José L.
Ortega-Alarcón, David
Rizzuti, Bruno
Palomino-Schätzlein, M.
Velázquez-Campoy, Adrián
Falcó, Alberto
author_role author
author2 Ortega-Alarcón, David
Rizzuti, Bruno
Palomino-Schätzlein, M.
Velázquez-Campoy, Adrián
Falcó, Alberto
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
Generalitat Valenciana
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Binding
Circular dichroism
Peptides
Isothermal titration calorimetry
NMR
Fluorescence
topic Binding
Circular dichroism
Peptides
Isothermal titration calorimetry
NMR
Fluorescence
description 18 pags., 4 figs., 5 tabs. -- This article belongs to the Special Issue Folding and Design of α-Helical Proteins and Peptides: Theory Meets Nanomaterials, Biotechnology and Health
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
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/10261/256259
url http://hdl.handle.net/10261/256259
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097991-B-I00
info:eu-repo/grantAgreement/MINECO//BFU2016-78232-P
http://dx.doi.org/10.3390/ijms221910805
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
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score 15,811543

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