CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase

Pyruvate carboxylase (PC) is a tetrameric enzyme that contains two active sites per subunit that catalyze two consecutive reactions. A mobile domain with an attached prosthetic biotin links both reactions, an initial biotin carboxylation and the subsequent carboxyl transfer to pyruvate substrate to...

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Autores: López-Alonso, Jorge P., Lázaro, Melisa, Gil-Cartón, David, Choi, Philip H., Dodu, Alexandra, Tong, Liang, Valle, Mikel
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/296774
Acceso en línea:http://hdl.handle.net/10261/296774
Access Level:acceso abierto
Palabra clave:Cryoelectron microscopy
Enzyme mechanisms
Transferases
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spelling CryoEM structural exploration of catalytically active enzyme pyruvate carboxylaseLópez-Alonso, Jorge P.Lázaro, MelisaGil-Cartón, DavidChoi, Philip H.Dodu, AlexandraTong, LiangValle, MikelCryoelectron microscopyEnzyme mechanismsTransferasesPyruvate carboxylase (PC) is a tetrameric enzyme that contains two active sites per subunit that catalyze two consecutive reactions. A mobile domain with an attached prosthetic biotin links both reactions, an initial biotin carboxylation and the subsequent carboxyl transfer to pyruvate substrate to produce oxaloacetate. Reaction sites are at long distance, and there are several co-factors that play as allosteric regulators. Here, using cryoEM we explore the structure of active PC tetramers focusing on active sites and on the conformational space of the oligomers. The results capture the mobile domain at both active sites and expose catalytic steps of both reactions at high resolution, allowing the identification of substrates and products. The analysis of catalytically active PC tetramers reveals the role of certain motions during enzyme functioning, and the structural changes in the presence of additional cofactors expose the mechanism for allosteric regulation.This study was supported by grants from the HFSP (RGP0062), and from the Spanish Ministerio de Ciencia e Innovación (PGC2018-098996-B-100) to M.V., and grants from the NIGMS (R35GM118093) and the NIAID (R01AI116669) to L.T.Peer reviewedNature Publishing GroupHuman Frontier Science ProgramMinisterio de Ciencia, Innovación y Universidades (España)Agencia Estatal de Investigación (España)National Institute of General Medical Sciences (US)National Institute of Allergy and Infectious Diseases (US)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202320232022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/296774reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PGC2018-098996-B-I00López-Alonso, Jorge P.; Lázaro, Melisa; Gil, David; Choi, Philip H. Dodu, Alexandra; Tong, Liang; Valle, Mikel. Correction: CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase. https://doi.org/10.1038/s41467-022-34543-8. http://hdl.handle.net/10261/296780https://doi.org/10.1038/s41467-022-33987-2Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2967742026-05-22T06:33:51Z
dc.title.none.fl_str_mv CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
title CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
spellingShingle CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
López-Alonso, Jorge P.
Cryoelectron microscopy
Enzyme mechanisms
Transferases
title_short CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
title_full CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
title_fullStr CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
title_full_unstemmed CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
title_sort CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase
dc.creator.none.fl_str_mv López-Alonso, Jorge P.
Lázaro, Melisa
Gil-Cartón, David
Choi, Philip H.
Dodu, Alexandra
Tong, Liang
Valle, Mikel
author López-Alonso, Jorge P.
author_facet López-Alonso, Jorge P.
Lázaro, Melisa
Gil-Cartón, David
Choi, Philip H.
Dodu, Alexandra
Tong, Liang
Valle, Mikel
author_role author
author2 Lázaro, Melisa
Gil-Cartón, David
Choi, Philip H.
Dodu, Alexandra
Tong, Liang
Valle, Mikel
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Human Frontier Science Program
Ministerio de Ciencia, Innovación y Universidades (España)
Agencia Estatal de Investigación (España)
National Institute of General Medical Sciences (US)
National Institute of Allergy and Infectious Diseases (US)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Cryoelectron microscopy
Enzyme mechanisms
Transferases
topic Cryoelectron microscopy
Enzyme mechanisms
Transferases
description Pyruvate carboxylase (PC) is a tetrameric enzyme that contains two active sites per subunit that catalyze two consecutive reactions. A mobile domain with an attached prosthetic biotin links both reactions, an initial biotin carboxylation and the subsequent carboxyl transfer to pyruvate substrate to produce oxaloacetate. Reaction sites are at long distance, and there are several co-factors that play as allosteric regulators. Here, using cryoEM we explore the structure of active PC tetramers focusing on active sites and on the conformational space of the oligomers. The results capture the mobile domain at both active sites and expose catalytic steps of both reactions at high resolution, allowing the identification of substrates and products. The analysis of catalytically active PC tetramers reveals the role of certain motions during enzyme functioning, and the structural changes in the presence of additional cofactors expose the mechanism for allosteric regulation.
publishDate 2022
dc.date.none.fl_str_mv 2022
2023
2023
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/10261/296774
url http://hdl.handle.net/10261/296774
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/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PGC2018-098996-B-I00
López-Alonso, Jorge P.; Lázaro, Melisa; Gil, David; Choi, Philip H. Dodu, Alexandra; Tong, Liang; Valle, Mikel. Correction: CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase. https://doi.org/10.1038/s41467-022-34543-8. http://hdl.handle.net/10261/296780
https://doi.org/10.1038/s41467-022-33987-2
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
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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