Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors

There exist two isoforms of cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) in pig populations that differ in a single amino acid (Met139Leu). The isoenzymes have different kinetic properties, affecting more strongly the Km and Vmax of nucleotides. They are associated to different phenotypes m...

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Detalhes bibliográficos
Autores: Hidalgo, Jorge, Latorre-Muro, Pedro, Carrodeguas, José A., Velázquez-Campoy, Adrián, Sancho, Javier, López-Buesa, P.
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2016
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/142015
Acesso em linha:http://hdl.handle.net/10261/142015
Access Level:Acceso aberto
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spelling Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitorsHidalgo, JorgeLatorre-Muro, PedroCarrodeguas, José A.Velázquez-Campoy, AdriánSancho, JavierLópez-Buesa, P.There exist two isoforms of cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) in pig populations that differ in a single amino acid (Met139Leu). The isoenzymes have different kinetic properties, affecting more strongly the Km and Vmax of nucleotides. They are associated to different phenotypes modifying traits of considerable economic interest. In this work we use inhibitors of phosphoenolpyruvate carboxykinase activity to search for further differences between these isoenzymes. On the one hand we have used the wellknown inhibitor 3-mercaptopicolinic acid. Its inhibition patterns were the same for both isoenzymes: a three-fold decrease of the Ki values for GTP in 139Met and 139Leu (273 and 873 μM, respectively). On the other hand, through screening of a chemical library we have found two novel compounds with inhibitory effects of a similar magnitude to that of 3-mercaptopicolinic acid but with less solubility and specificity. One of these novel compounds, (N'1-({5-[1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl]-2-thienyl}methylidene)-2,4-dichlorobenzene-1-carbohydrazide), exhibited significantly different inhibitory effects on either isoenzyme: it enhanced threefold the apparent Km value for GTP in 139Met, whereas in139Leu, it reduced it from 99 to 69 μM. The finding of those significant differences in the binding of GTP reinforces the hypothesis that the Met139Leu substitution affects strongly the nucleotide binding site of PEPCK-C.Supported by research grants AGL2008-01487ALI (www.mineco.gob.es), DGA-IAF FITE2012/2013 (www.aragob.es), and UZ2014-CIE-03 (www.unizar.es) to P.L.B., AGL2015-66177 to P.L.B. and J.A.C., and grants BFU2013-47064-P (www.micinn.es), BIO2014-57314-REDT (www.mineco.gob.es) and PI078/08 to J.S. P.L.Peer ReviewedPublic Library of ScienceMinisterio de Economía y Competitividad (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2017201720162017info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/142015reponame: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/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/AGL2015-66177-Rinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2013-47064-Pinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2014-57314-REDTSíinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1420152026-05-22T06:33:51Z
dc.title.none.fl_str_mv Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
title Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
spellingShingle Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
Hidalgo, Jorge
title_short Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
title_full Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
title_fullStr Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
title_full_unstemmed Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
title_sort Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors
dc.creator.none.fl_str_mv Hidalgo, Jorge
Latorre-Muro, Pedro
Carrodeguas, José A.
Velázquez-Campoy, Adrián
Sancho, Javier
López-Buesa, P.
author Hidalgo, Jorge
author_facet Hidalgo, Jorge
Latorre-Muro, Pedro
Carrodeguas, José A.
Velázquez-Campoy, Adrián
Sancho, Javier
López-Buesa, P.
author_role author
author2 Latorre-Muro, Pedro
Carrodeguas, José A.
Velázquez-Campoy, Adrián
Sancho, Javier
López-Buesa, P.
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
description There exist two isoforms of cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) in pig populations that differ in a single amino acid (Met139Leu). The isoenzymes have different kinetic properties, affecting more strongly the Km and Vmax of nucleotides. They are associated to different phenotypes modifying traits of considerable economic interest. In this work we use inhibitors of phosphoenolpyruvate carboxykinase activity to search for further differences between these isoenzymes. On the one hand we have used the wellknown inhibitor 3-mercaptopicolinic acid. Its inhibition patterns were the same for both isoenzymes: a three-fold decrease of the Ki values for GTP in 139Met and 139Leu (273 and 873 μM, respectively). On the other hand, through screening of a chemical library we have found two novel compounds with inhibitory effects of a similar magnitude to that of 3-mercaptopicolinic acid but with less solubility and specificity. One of these novel compounds, (N'1-({5-[1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl]-2-thienyl}methylidene)-2,4-dichlorobenzene-1-carbohydrazide), exhibited significantly different inhibitory effects on either isoenzyme: it enhanced threefold the apparent Km value for GTP in 139Met, whereas in139Leu, it reduced it from 99 to 69 μM. The finding of those significant differences in the binding of GTP reinforces the hypothesis that the Met139Leu substitution affects strongly the nucleotide binding site of PEPCK-C.
publishDate 2016
dc.date.none.fl_str_mv 2016
2017
2017
2017
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/10261/142015
url http://hdl.handle.net/10261/142015
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
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#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/AGL2015-66177-R
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2013-47064-P
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2014-57314-REDT
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
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
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