On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli

Autores
Ebrecht, Ana Cristina; Orlof, Agnieszka M.; Sasoni, Natalia; Figueroa, Carlos Maria; Iglesias, Alberto Alvaro; Ballicora, Miguel A.
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In bacteria, UDP-glucose is a central intermediate in carbohydrate metabolism. The enzyme responsible for its synthesis is encoded by the galU gene and its deletion generates cells unable to ferment galactose. In some bacteria, there is a second gene, galF, encoding for a protein with high sequence identity to GalU. However, the role of GalF has been contradictory regarding its catalytic capability and not well understood. In this work we show that GalF derives from a catalytic (UDP-glucose pyrophosphorylase) ancestor, but its activity is very low compared to GalU. We demonstrated that GalF has some residual UDP-glucose pyrophosphorylase activity by in vitro and in vivo experiments in which the phenotype of a galU- strain was reverted by the over-expression of GalF and its mutant. To demonstrate its evolutionary path of "enzyme inactivation" we enhanced the catalysis by mutagenesis and showed the importance of the quaternary structure. This study provides important information to understand the structural and functional evolutionary origin of the protein GalF in enteric bacteria.
Fil: Ebrecht, Ana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina. Loyola University; Estados Unidos
Fil: Orlof, Agnieszka M.. Loyola University; Estados Unidos
Fil: Sasoni, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Figueroa, Carlos Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Ballicora, Miguel A.. Loyola University; Estados Unidos
Materia
Catalytic Residues
Enzyme Evolution
Enzyme Inactivation
Enzyme Resurrection
Galactose Metabolism
Nucleotidyltransferase
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/39726
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/39726
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coliEbrecht, Ana CristinaOrlof, Agnieszka M.Sasoni, NataliaFigueroa, Carlos MariaIglesias, Alberto AlvaroBallicora, Miguel A.Catalytic ResiduesEnzyme EvolutionEnzyme InactivationEnzyme ResurrectionGalactose MetabolismNucleotidyltransferasehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1In bacteria, UDP-glucose is a central intermediate in carbohydrate metabolism. The enzyme responsible for its synthesis is encoded by the galU gene and its deletion generates cells unable to ferment galactose. In some bacteria, there is a second gene, galF, encoding for a protein with high sequence identity to GalU. However, the role of GalF has been contradictory regarding its catalytic capability and not well understood. In this work we show that GalF derives from a catalytic (UDP-glucose pyrophosphorylase) ancestor, but its activity is very low compared to GalU. We demonstrated that GalF has some residual UDP-glucose pyrophosphorylase activity by in vitro and in vivo experiments in which the phenotype of a galU- strain was reverted by the over-expression of GalF and its mutant. To demonstrate its evolutionary path of "enzyme inactivation" we enhanced the catalysis by mutagenesis and showed the importance of the quaternary structure. This study provides important information to understand the structural and functional evolutionary origin of the protein GalF in enteric bacteria.Fil: Ebrecht, Ana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina. Loyola University; Estados UnidosFil: Orlof, Agnieszka M.. Loyola University; Estados UnidosFil: Sasoni, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Figueroa, Carlos Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Ballicora, Miguel A.. Loyola University; Estados UnidosFrontiers Research Foundation2015-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/39726Ebrecht, Ana Cristina; Orlof, Agnieszka M.; Sasoni, Natalia; Figueroa, Carlos Maria; Iglesias, Alberto Alvaro; et al.; On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli; Frontiers Research Foundation; Frontiers in Microbiology; 6; 1253; 11-2015; 1-131664-302XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fmicb.2015.01253/fullinfo:eu-repo/semantics/altIdentifier/doi/10.3389/fmicb.2015.01253info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-02-06T13:18:15Zoai:ri.conicet.gov.ar:11336/39726instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982026-02-06 13:18:15.988CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
title On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
spellingShingle On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
Ebrecht, Ana Cristina
Catalytic Residues
Enzyme Evolution
Enzyme Inactivation
Enzyme Resurrection
Galactose Metabolism
Nucleotidyltransferase
title_short On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
title_full On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
title_fullStr On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
title_full_unstemmed On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
title_sort On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli
dc.creator.none.fl_str_mv Ebrecht, Ana Cristina
Orlof, Agnieszka M.
Sasoni, Natalia
Figueroa, Carlos Maria
Iglesias, Alberto Alvaro
Ballicora, Miguel A.
author Ebrecht, Ana Cristina
author_facet Ebrecht, Ana Cristina
Orlof, Agnieszka M.
Sasoni, Natalia
Figueroa, Carlos Maria
Iglesias, Alberto Alvaro
Ballicora, Miguel A.
author_role author
author2 Orlof, Agnieszka M.
Sasoni, Natalia
Figueroa, Carlos Maria
Iglesias, Alberto Alvaro
Ballicora, Miguel A.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Catalytic Residues
Enzyme Evolution
Enzyme Inactivation
Enzyme Resurrection
Galactose Metabolism
Nucleotidyltransferase
topic Catalytic Residues
Enzyme Evolution
Enzyme Inactivation
Enzyme Resurrection
Galactose Metabolism
Nucleotidyltransferase
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In bacteria, UDP-glucose is a central intermediate in carbohydrate metabolism. The enzyme responsible for its synthesis is encoded by the galU gene and its deletion generates cells unable to ferment galactose. In some bacteria, there is a second gene, galF, encoding for a protein with high sequence identity to GalU. However, the role of GalF has been contradictory regarding its catalytic capability and not well understood. In this work we show that GalF derives from a catalytic (UDP-glucose pyrophosphorylase) ancestor, but its activity is very low compared to GalU. We demonstrated that GalF has some residual UDP-glucose pyrophosphorylase activity by in vitro and in vivo experiments in which the phenotype of a galU- strain was reverted by the over-expression of GalF and its mutant. To demonstrate its evolutionary path of "enzyme inactivation" we enhanced the catalysis by mutagenesis and showed the importance of the quaternary structure. This study provides important information to understand the structural and functional evolutionary origin of the protein GalF in enteric bacteria.
Fil: Ebrecht, Ana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina. Loyola University; Estados Unidos
Fil: Orlof, Agnieszka M.. Loyola University; Estados Unidos
Fil: Sasoni, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Figueroa, Carlos Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Ballicora, Miguel A.. Loyola University; Estados Unidos
description In bacteria, UDP-glucose is a central intermediate in carbohydrate metabolism. The enzyme responsible for its synthesis is encoded by the galU gene and its deletion generates cells unable to ferment galactose. In some bacteria, there is a second gene, galF, encoding for a protein with high sequence identity to GalU. However, the role of GalF has been contradictory regarding its catalytic capability and not well understood. In this work we show that GalF derives from a catalytic (UDP-glucose pyrophosphorylase) ancestor, but its activity is very low compared to GalU. We demonstrated that GalF has some residual UDP-glucose pyrophosphorylase activity by in vitro and in vivo experiments in which the phenotype of a galU- strain was reverted by the over-expression of GalF and its mutant. To demonstrate its evolutionary path of "enzyme inactivation" we enhanced the catalysis by mutagenesis and showed the importance of the quaternary structure. This study provides important information to understand the structural and functional evolutionary origin of the protein GalF in enteric bacteria.
publishDate 2015
dc.date.none.fl_str_mv 2015-11
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/39726
Ebrecht, Ana Cristina; Orlof, Agnieszka M.; Sasoni, Natalia; Figueroa, Carlos Maria; Iglesias, Alberto Alvaro; et al.; On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli; Frontiers Research Foundation; Frontiers in Microbiology; 6; 1253; 11-2015; 1-13
1664-302X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/39726
identifier_str_mv Ebrecht, Ana Cristina; Orlof, Agnieszka M.; Sasoni, Natalia; Figueroa, Carlos Maria; Iglesias, Alberto Alvaro; et al.; On the ancestral UDP-glucose pyrophosphorylase activity of GalF from Escherichia coli; Frontiers Research Foundation; Frontiers in Microbiology; 6; 1253; 11-2015; 1-13
1664-302X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fmicb.2015.01253/full
info:eu-repo/semantics/altIdentifier/doi/10.3389/fmicb.2015.01253
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Frontiers Research Foundation
publisher.none.fl_str_mv Frontiers Research Foundation
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.595271

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