Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis

[EN]ADP-dependent glucokinases represent a unique family of kinases that belong to the ribokinase superfamily, being present mainly in hyperthermophilic archaea. For these enzymes there is no agreement about the magnitude of the structural transitions associated with ligand binding and whether they...

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Autores: Rivas-Pardo, J. A., Herrera-Morande, A., Castro-Fernández, V., Fernández, Francisco J., Vega, María Cristina, Guixé, V.
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2013
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/95792
Acesso em linha:http://hdl.handle.net/10261/95792
Access Level:acceso abierto
Palavra-chave:Biocatalysis
Crystal structures
Enzyme kinetics
Enzyme structure
Enzymes
Glucose
Protein interactions
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spelling Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for CatalysisRivas-Pardo, J. A.Herrera-Morande, A.Castro-Fernández, V.Fernández, Francisco J.Vega, María CristinaGuixé, V.BiocatalysisCrystal structuresEnzyme kineticsEnzyme structureEnzymesGlucoseProtein interactions[EN]ADP-dependent glucokinases represent a unique family of kinases that belong to the ribokinase superfamily, being present mainly in hyperthermophilic archaea. For these enzymes there is no agreement about the magnitude of the structural transitions associated with ligand binding and whether they are meaningful to the function of the enzyme. We used the ADP-dependent glucokinase from Termococcus litoralis as a model to investigate the conformational changes observed in X-ray crystallographic structures upon substrate binding and to compare them with those determined in solution in order to understand their interplay with the glucokinase function. Initial velocity studies indicate that catalysis follows a sequential ordered mechanism that correlates with the structural transitions experienced by the enzyme in solution and in the crystal state. The combined data allowed us to resolve the open-closed conformational transition that accounts for the complete reaction cycle and to identify the corresponding clusters of aminoacids residues responsible for it. These results provide molecular bases for a general mechanism conserved across the ADP-dependent kinase familyThis work was supported by Fondo Nacional de Desarrollo Científico y Tecnológico FONDECYT grant 1110137 to V.G, Spanish Ministry of Science and Innovation grants PET2008_0101, BIO2009-11184 and BFU2010-22260-C02-02 and the EC project ComplexINC (Framework Programme 7 (FP7) under grant agreement no. 279039) to M.C.V. The SAXS measurements (SAXS1 and SAXS2 beamlines) were supported by LNLS (Laboratorio Nacional de Luz Sincrotron), Sao Paulo, Brazil, to V.G. and J.A.R-P. We also thank the Departamento de Postgrado y Postitulo, Universidad de Chile by the support given to J.A.R-P to accomplish the SAXS studies in Sao Paulo. J.A.R-P and V.C-F are PhD fellowship from Comisión Nacional de Investigación Científica y Tecnológica de Chile. A.H-M. acknowledges the support of the PhD program in Molecular Biotechnology of the Universitat de Barcelona.Peer reviewedPublic Library of Science201420142013info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/95792reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#279039http://dx.doi.org/10.1371/journal.pone.0066687info:eu-repo/semantics/openAccessoai:digital.csic.es:10261/957922026-05-22T06:33:51Z
dc.title.none.fl_str_mv Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
title Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
spellingShingle Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
Rivas-Pardo, J. A.
Biocatalysis
Crystal structures
Enzyme kinetics
Enzyme structure
Enzymes
Glucose
Protein interactions
title_short Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
title_full Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
title_fullStr Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
title_full_unstemmed Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
title_sort Crystal Structure, SAXS and Kinetic Mechanism of Hyperthermophilic ADP-Dependent Glucokinase from Thermococcus litoralis Reveal a Conserved Mechanism for Catalysis
dc.creator.none.fl_str_mv Rivas-Pardo, J. A.
Herrera-Morande, A.
Castro-Fernández, V.
Fernández, Francisco J.
Vega, María Cristina
Guixé, V.
author Rivas-Pardo, J. A.
author_facet Rivas-Pardo, J. A.
Herrera-Morande, A.
Castro-Fernández, V.
Fernández, Francisco J.
Vega, María Cristina
Guixé, V.
author_role author
author2 Herrera-Morande, A.
Castro-Fernández, V.
Fernández, Francisco J.
Vega, María Cristina
Guixé, V.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Biocatalysis
Crystal structures
Enzyme kinetics
Enzyme structure
Enzymes
Glucose
Protein interactions
topic Biocatalysis
Crystal structures
Enzyme kinetics
Enzyme structure
Enzymes
Glucose
Protein interactions
description [EN]ADP-dependent glucokinases represent a unique family of kinases that belong to the ribokinase superfamily, being present mainly in hyperthermophilic archaea. For these enzymes there is no agreement about the magnitude of the structural transitions associated with ligand binding and whether they are meaningful to the function of the enzyme. We used the ADP-dependent glucokinase from Termococcus litoralis as a model to investigate the conformational changes observed in X-ray crystallographic structures upon substrate binding and to compare them with those determined in solution in order to understand their interplay with the glucokinase function. Initial velocity studies indicate that catalysis follows a sequential ordered mechanism that correlates with the structural transitions experienced by the enzyme in solution and in the crystal state. The combined data allowed us to resolve the open-closed conformational transition that accounts for the complete reaction cycle and to identify the corresponding clusters of aminoacids residues responsible for it. These results provide molecular bases for a general mechanism conserved across the ADP-dependent kinase family
publishDate 2013
dc.date.none.fl_str_mv 2013
2014
2014
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/10261/95792
url http://hdl.handle.net/10261/95792
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
279039
http://dx.doi.org/10.1371/journal.pone.0066687
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
repository.name.fl_str_mv
repository.mail.fl_str_mv
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