Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase

Autores
Soares, Jose Sergio M.; Gentile, Agustina; Scorsato, Valeria; Lima, Aline da C.; Kiyota, Eduardo; Santos, Marcelo Leite Dos; Piattoni, Claudia Vanesa; Huber, Steven C.; Aparicio, Ricardo; Menossi, Marcelo
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Sugarcane is a monocot plant that accumulates sucrose to levels of up to 50% of dry weight in the stalk. The mechanisms that are involved in sucrose accumulation in sugarcane are not well understood, and little is known with regard to factors that control the extent of sucrose storage in the stalks. UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) is an enzyme that produces UDP-glucose, a key precursor for sucrose metabolism and cell wall biosynthesis. The objective of this work was to gain insights into the ScUGPase-1 expression pattern and regulatory mechanisms that control protein activity. ScUGPase-1 expression was negatively correlated with the sucrose content in the internodes during development, and only slight differences in the expression patterns were observed between two cultivars that differ in sucrose content. The intracellular localization of ScUGPase-1 indicated partial membrane association of this soluble protein in both the leaves and internodes. Using a phospho-specific antibody, we observed that ScUGPase-1 was phosphorylated in vivo at the Ser-419 site in the soluble and membrane fractions from the leaves but not from the internodes. The purified recombinant enzyme was kinetically characterized in the direction of UDP-glucose formation, and the enzyme activity was affected by redox modification. Preincubation with H2O2 strongly inhibited this activity, which could be reversed by DTT. Small angle x-ray scattering analysis indicated that the dimer interface is located at the C terminus and provided the first structural model of the dimer of sugarcane UGPase in solution.
Fil: Soares, Jose Sergio M.. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; Brasil
Fil: Gentile, Agustina. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; Brasil
Fil: Scorsato, Valeria. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Lima, Aline da C.. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Kiyota, Eduardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Santos, Marcelo Leite Dos . Universidade Federal do Sergipe. Centro de Ciências Exatas e Tecnologia. Núcleo de Química; Brasil
Fil: Piattoni, Claudia Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina. Universidad Nacional del Litoral; Argentina
Fil: Huber, Steven C.. University of Illinois at Urbana-Champaign. Department of Agriculture Agricultural Research Service, and Department of Plant Biology; Estados Unidos
Fil: Aparicio, Ricardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Menossi, Marcelo. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; Brasil
Materia
redox regulation
kinetics
small-angle X-ray scattering (SAXS)
gene expression
protein phosphorylation
sucrose
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/14172
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/14172
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose PyrophosphorylaseSoares, Jose Sergio M.Gentile, AgustinaScorsato, ValeriaLima, Aline da C.Kiyota, EduardoSantos, Marcelo Leite Dos Piattoni, Claudia VanesaHuber, Steven C.Aparicio, RicardoMenossi, Marceloredox regulationkineticssmall-angle X-ray scattering (SAXS)gene expressionprotein phosphorylationsucrosehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Sugarcane is a monocot plant that accumulates sucrose to levels of up to 50% of dry weight in the stalk. The mechanisms that are involved in sucrose accumulation in sugarcane are not well understood, and little is known with regard to factors that control the extent of sucrose storage in the stalks. UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) is an enzyme that produces UDP-glucose, a key precursor for sucrose metabolism and cell wall biosynthesis. The objective of this work was to gain insights into the ScUGPase-1 expression pattern and regulatory mechanisms that control protein activity. ScUGPase-1 expression was negatively correlated with the sucrose content in the internodes during development, and only slight differences in the expression patterns were observed between two cultivars that differ in sucrose content. The intracellular localization of ScUGPase-1 indicated partial membrane association of this soluble protein in both the leaves and internodes. Using a phospho-specific antibody, we observed that ScUGPase-1 was phosphorylated in vivo at the Ser-419 site in the soluble and membrane fractions from the leaves but not from the internodes. The purified recombinant enzyme was kinetically characterized in the direction of UDP-glucose formation, and the enzyme activity was affected by redox modification. Preincubation with H2O2 strongly inhibited this activity, which could be reversed by DTT. Small angle x-ray scattering analysis indicated that the dimer interface is located at the C terminus and provided the first structural model of the dimer of sugarcane UGPase in solution.Fil: Soares, Jose Sergio M.. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; BrasilFil: Gentile, Agustina. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; BrasilFil: Scorsato, Valeria. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Lima, Aline da C.. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Kiyota, Eduardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Santos, Marcelo Leite Dos . Universidade Federal do Sergipe. Centro de Ciências Exatas e Tecnologia. Núcleo de Química; BrasilFil: Piattoni, Claudia Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina. Universidad Nacional del Litoral; ArgentinaFil: Huber, Steven C.. University of Illinois at Urbana-Champaign. Department of Agriculture Agricultural Research Service, and Department of Plant Biology; Estados UnidosFil: Aparicio, Ricardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; BrasilFil: Menossi, Marcelo. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; BrasilAmerican Society for Biochemistry and Molecular Biology2014-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/14172Soares, Jose Sergio M.; Gentile, Agustina; Scorsato, Valeria; Lima, Aline da C.; Kiyota, Eduardo; et al.; Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase; American Society for Biochemistry and Molecular Biology; Journal Of Biological Chemistry (online); 289; 11-2014; 33364-333770021-92581083-351Xenginfo:eu-repo/semantics/altIdentifier/url/http://dx.doi.org//10.1074/jbc.M114.590125info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/289/48/33364info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-01-08T12:51:48Zoai:ri.conicet.gov.ar:11336/14172instacron: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:34982026-01-08 12:51:49.208CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
title Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
spellingShingle Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
Soares, Jose Sergio M.
redox regulation
kinetics
small-angle X-ray scattering (SAXS)
gene expression
protein phosphorylation
sucrose
title_short Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
title_full Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
title_fullStr Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
title_full_unstemmed Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
title_sort Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase
dc.creator.none.fl_str_mv Soares, Jose Sergio M.
Gentile, Agustina
Scorsato, Valeria
Lima, Aline da C.
Kiyota, Eduardo
Santos, Marcelo Leite Dos
Piattoni, Claudia Vanesa
Huber, Steven C.
Aparicio, Ricardo
Menossi, Marcelo
author Soares, Jose Sergio M.
author_facet Soares, Jose Sergio M.
Gentile, Agustina
Scorsato, Valeria
Lima, Aline da C.
Kiyota, Eduardo
Santos, Marcelo Leite Dos
Piattoni, Claudia Vanesa
Huber, Steven C.
Aparicio, Ricardo
Menossi, Marcelo
author_role author
author2 Gentile, Agustina
Scorsato, Valeria
Lima, Aline da C.
Kiyota, Eduardo
Santos, Marcelo Leite Dos
Piattoni, Claudia Vanesa
Huber, Steven C.
Aparicio, Ricardo
Menossi, Marcelo
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv redox regulation
kinetics
small-angle X-ray scattering (SAXS)
gene expression
protein phosphorylation
sucrose
topic redox regulation
kinetics
small-angle X-ray scattering (SAXS)
gene expression
protein phosphorylation
sucrose
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Sugarcane is a monocot plant that accumulates sucrose to levels of up to 50% of dry weight in the stalk. The mechanisms that are involved in sucrose accumulation in sugarcane are not well understood, and little is known with regard to factors that control the extent of sucrose storage in the stalks. UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) is an enzyme that produces UDP-glucose, a key precursor for sucrose metabolism and cell wall biosynthesis. The objective of this work was to gain insights into the ScUGPase-1 expression pattern and regulatory mechanisms that control protein activity. ScUGPase-1 expression was negatively correlated with the sucrose content in the internodes during development, and only slight differences in the expression patterns were observed between two cultivars that differ in sucrose content. The intracellular localization of ScUGPase-1 indicated partial membrane association of this soluble protein in both the leaves and internodes. Using a phospho-specific antibody, we observed that ScUGPase-1 was phosphorylated in vivo at the Ser-419 site in the soluble and membrane fractions from the leaves but not from the internodes. The purified recombinant enzyme was kinetically characterized in the direction of UDP-glucose formation, and the enzyme activity was affected by redox modification. Preincubation with H2O2 strongly inhibited this activity, which could be reversed by DTT. Small angle x-ray scattering analysis indicated that the dimer interface is located at the C terminus and provided the first structural model of the dimer of sugarcane UGPase in solution.
Fil: Soares, Jose Sergio M.. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; Brasil
Fil: Gentile, Agustina. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; Brasil
Fil: Scorsato, Valeria. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Lima, Aline da C.. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Kiyota, Eduardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Santos, Marcelo Leite Dos . Universidade Federal do Sergipe. Centro de Ciências Exatas e Tecnologia. Núcleo de Química; Brasil
Fil: Piattoni, Claudia Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina. Universidad Nacional del Litoral; Argentina
Fil: Huber, Steven C.. University of Illinois at Urbana-Champaign. Department of Agriculture Agricultural Research Service, and Department of Plant Biology; Estados Unidos
Fil: Aparicio, Ricardo. Universidade Estadual de Campinas. Instituto de Química. Laboratório de Biologia Estrutural e Cristalografia; Brasil
Fil: Menossi, Marcelo. Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Genética, Evolução e Bioagentes; Brasil
description Sugarcane is a monocot plant that accumulates sucrose to levels of up to 50% of dry weight in the stalk. The mechanisms that are involved in sucrose accumulation in sugarcane are not well understood, and little is known with regard to factors that control the extent of sucrose storage in the stalks. UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) is an enzyme that produces UDP-glucose, a key precursor for sucrose metabolism and cell wall biosynthesis. The objective of this work was to gain insights into the ScUGPase-1 expression pattern and regulatory mechanisms that control protein activity. ScUGPase-1 expression was negatively correlated with the sucrose content in the internodes during development, and only slight differences in the expression patterns were observed between two cultivars that differ in sucrose content. The intracellular localization of ScUGPase-1 indicated partial membrane association of this soluble protein in both the leaves and internodes. Using a phospho-specific antibody, we observed that ScUGPase-1 was phosphorylated in vivo at the Ser-419 site in the soluble and membrane fractions from the leaves but not from the internodes. The purified recombinant enzyme was kinetically characterized in the direction of UDP-glucose formation, and the enzyme activity was affected by redox modification. Preincubation with H2O2 strongly inhibited this activity, which could be reversed by DTT. Small angle x-ray scattering analysis indicated that the dimer interface is located at the C terminus and provided the first structural model of the dimer of sugarcane UGPase in solution.
publishDate 2014
dc.date.none.fl_str_mv 2014-11
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/11336/14172
Soares, Jose Sergio M.; Gentile, Agustina; Scorsato, Valeria; Lima, Aline da C.; Kiyota, Eduardo; et al.; Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase; American Society for Biochemistry and Molecular Biology; Journal Of Biological Chemistry (online); 289; 11-2014; 33364-33377
0021-9258
1083-351X
url http://hdl.handle.net/11336/14172
identifier_str_mv Soares, Jose Sergio M.; Gentile, Agustina; Scorsato, Valeria; Lima, Aline da C.; Kiyota, Eduardo; et al.; Oligomerization, Membrane Association, and in Vivo Phosphorylation of Sugarcane UDP-glucose Pyrophosphorylase; American Society for Biochemistry and Molecular Biology; Journal Of Biological Chemistry (online); 289; 11-2014; 33364-33377
0021-9258
1083-351X
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://dx.doi.org//10.1074/jbc.M114.590125
info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/289/48/33364
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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 13.25844

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