The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability

23 pags., 5 figs., 3 tabs. -- This article belongs to the Special Issue Structural, Functional and Folding Strategies of Oligomeric Proteins

Detalhes bibliográficos
Autores: Neira, José L., Cámara-Artigas, Ana, Hernández-Cifre, José G., Ortore, M.G.
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:10261/253353
Acesso em linha:http://hdl.handle.net/10261/253353
Access Level:acceso abierto
Palavra-chave:Circular dichroism
Conformational stability
Fluorescence
Phosphorylation
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spelling The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stabilityNeira, José L.Cámara-Artigas, AnaHernández-Cifre, José G.Ortore, M.G.Circular dichroismConformational stabilityFluorescencePhosphorylation23 pags., 5 figs., 3 tabs. -- This article belongs to the Special Issue Structural, Functional and Folding Strategies of Oligomeric ProteinsThe histidine phosphocarrier protein (HPr) kinase/phosphorylase (HPrK/P) modulates the phosphorylation state of the HPr protein, and it is involved in the use of carbon sources by Gram-positive bacteria. Its X-ray structure, as concluded from crystals of proteins from several species, is a hexamer; however, there are no studies about its conformational stability, and how its structure is modified by the pH. We have embarked on the conformational characterization of HPrK/P of Bacillus subtilis (bsHPrK/P) in solution by using several spectroscopic (namely, fluorescence and circular dichroism (CD)) and biophysical techniques (namely, small-angle X-ray-scattering (SAXS) and dynamic light-scattering (DLS)). bsHPrK/P was mainly a hexamer in solution at pH 7.0, in the presence of phosphate. The protein had a high conformational stability, with an apparent thermal denaturation midpoint of ~70 °C, at pH 7.0, as monitored by fluorescence and CD. The protein was very pH-sensitive, precipitated between pH 3.5 and 6.5; below pH 3.5, it had a molten-globule-like conformation; and it acquired a native-like structure in a narrow pH range (between pH 7.0 and 8.0). Guanidinium hydrochloride (GdmCl) denaturation occurred through an oligomeric intermediate. On the other hand, urea denaturation occurred as a single transition, in the range of concentrations between 1.8 and 18 µM, as detected by far-UV CD and fluorescence.This research was funded by Spanish Ministry of Economy and Competitiveness and European ERDF Funds (MCIU/AEI/FEDER, EU) [RTI2018-097991-B-I00 to JLN, BIO2016-78020-R to ACA and CTQ2017-85425-P to JGHC]; JGHC thanks Fundación Séneca, Región de Murcia for funding (20933/PI/18).Peer reviewedMultidisciplinary Digital Publishing InstituteMinisterio de Ciencia, Innovación y Universidades (España)Ministerio de Economía y Competitividad (España)Fundación SénecaConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202120212021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/253353reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##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/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2017-85425-Pinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-78020-Rhttps://doi.org/10.3390/ijms22063231Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2533532026-05-22T06:33:51Z
dc.title.none.fl_str_mv The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
title The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
spellingShingle The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
Neira, José L.
Circular dichroism
Conformational stability
Fluorescence
Phosphorylation
title_short The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
title_full The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
title_fullStr The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
title_full_unstemmed The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
title_sort The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability
dc.creator.none.fl_str_mv Neira, José L.
Cámara-Artigas, Ana
Hernández-Cifre, José G.
Ortore, M.G.
author Neira, José L.
author_facet Neira, José L.
Cámara-Artigas, Ana
Hernández-Cifre, José G.
Ortore, M.G.
author_role author
author2 Cámara-Artigas, Ana
Hernández-Cifre, José G.
Ortore, M.G.
author2_role author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Ministerio de Economía y Competitividad (España)
Fundación Séneca
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Circular dichroism
Conformational stability
Fluorescence
Phosphorylation
topic Circular dichroism
Conformational stability
Fluorescence
Phosphorylation
description 23 pags., 5 figs., 3 tabs. -- This article belongs to the Special Issue Structural, Functional and Folding Strategies of Oligomeric Proteins
publishDate 2021
dc.date.none.fl_str_mv 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/10261/253353
url http://hdl.handle.net/10261/253353
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#
#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/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2017-85425-P
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-78020-R
https://doi.org/10.3390/ijms22063231
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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