Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase

Autores
Hill, B. L.; Mascarenhas, R.; Patel, H. P.; Asención Diez, Matías Damián; Wu, R.; Iglesias, Alberto Alvaro; Liu, D.; Ballicora, M. A.
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The pathways for biosynthesis of glycogen inbacteria and starch in plants are evolutionarily andbiochemically related. They are regulated primarily by ADP?glucose pyrophosphorylase, which evolved to satisfy metabolic requirements of a particular organism. Despite the importance of these two pathways, little is known about the mechanism that controls pyrophosphorylase activity or the location of its allosteric sites. Here, we report pyruvate-bound crystal structures of ADP-glucose pyrophosphorylase from the bacterium Agrobacterium tumefaciens, identifying a previously elusive activator site for the enzyme. We found that the tetrameric enzyme binds two molecules of pyruvate in a planar conformation. Each binding site is located in a crevice between the C-terminal domains of two subunits where they stack via a distinct β-helix region. Pyruvate interacts with the side chain of Lys-43 and with the peptide backbone of Ser-328 and Gly-329 from both subunits. These structural insights led to the design of two variants with altered regulator properties. In one variant (K43A), the allosteric effect was absent, whereas in the other (G329D), the introduced Asp mimicked the presence of pyruvate. The latter generated an enzyme that was pre-activated and insensitive to further activation by pyruvate. Our study furnishes a deeper understanding of how glycogen biosynthesis is regulated in bacteria and the mechanism by which transgenic plants increased their starch production. These insights will facilitate rational approaches to enzyme engineering for starch production in crops of agricultural interest and will promote further study of allosteric signal transmission and molecular evolution in this important enzyme family.
Fil: Hill, B. L.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Mascarenhas, R.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Patel, H. P.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Asención Diez, Matías Damián. 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: Wu, R.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
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: Liu, D.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Ballicora, M. A.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Materia
GLUCOSE-1-PHOSPHATE ADENYLYLTRANSFERASE
GLYCOGEN BIOSYNTHESIS
STARCH BIOSYNTHESIS
ALLOSTERISM
ENZYME EVOLUTION
GLUCAN BIOSYNTHESIS
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:11336/118795
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/118795
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylaseHill, B. L.Mascarenhas, R.Patel, H. P.Asención Diez, Matías DamiánWu, R.Iglesias, Alberto AlvaroLiu, D.Ballicora, M. A.GLUCOSE-1-PHOSPHATE ADENYLYLTRANSFERASEGLYCOGEN BIOSYNTHESISSTARCH BIOSYNTHESISALLOSTERISMENZYME EVOLUTIONGLUCAN BIOSYNTHESIShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The pathways for biosynthesis of glycogen inbacteria and starch in plants are evolutionarily andbiochemically related. They are regulated primarily by ADP?glucose pyrophosphorylase, which evolved to satisfy metabolic requirements of a particular organism. Despite the importance of these two pathways, little is known about the mechanism that controls pyrophosphorylase activity or the location of its allosteric sites. Here, we report pyruvate-bound crystal structures of ADP-glucose pyrophosphorylase from the bacterium Agrobacterium tumefaciens, identifying a previously elusive activator site for the enzyme. We found that the tetrameric enzyme binds two molecules of pyruvate in a planar conformation. Each binding site is located in a crevice between the C-terminal domains of two subunits where they stack via a distinct β-helix region. Pyruvate interacts with the side chain of Lys-43 and with the peptide backbone of Ser-328 and Gly-329 from both subunits. These structural insights led to the design of two variants with altered regulator properties. In one variant (K43A), the allosteric effect was absent, whereas in the other (G329D), the introduced Asp mimicked the presence of pyruvate. The latter generated an enzyme that was pre-activated and insensitive to further activation by pyruvate. Our study furnishes a deeper understanding of how glycogen biosynthesis is regulated in bacteria and the mechanism by which transgenic plants increased their starch production. These insights will facilitate rational approaches to enzyme engineering for starch production in crops of agricultural interest and will promote further study of allosteric signal transmission and molecular evolution in this important enzyme family.Fil: Hill, B. L.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados UnidosFil: Mascarenhas, R.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados UnidosFil: Patel, H. P.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados UnidosFil: Asención Diez, Matías Damián. 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: Wu, R.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados UnidosFil: 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: Liu, D.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados UnidosFil: Ballicora, M. A.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados UnidosAmerican Society for Biochemistry and Molecular Biology2019-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/118795Hill, B. L.; Mascarenhas, R.; Patel, H. P.; Asención Diez, Matías Damián; Wu, R.; et al.; Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 294; 1-2019; 1338-13480021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/lookup/doi/10.1074/jbc.RA118.004246info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.RA118.004246info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T12:12:55Zoai:ri.conicet.gov.ar:11336/118795instacron: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:34982025-10-22 12:12:55.439CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
title Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
spellingShingle Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
Hill, B. L.
GLUCOSE-1-PHOSPHATE ADENYLYLTRANSFERASE
GLYCOGEN BIOSYNTHESIS
STARCH BIOSYNTHESIS
ALLOSTERISM
ENZYME EVOLUTION
GLUCAN BIOSYNTHESIS
title_short Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
title_full Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
title_fullStr Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
title_full_unstemmed Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
title_sort Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase
dc.creator.none.fl_str_mv Hill, B. L.
Mascarenhas, R.
Patel, H. P.
Asención Diez, Matías Damián
Wu, R.
Iglesias, Alberto Alvaro
Liu, D.
Ballicora, M. A.
author Hill, B. L.
author_facet Hill, B. L.
Mascarenhas, R.
Patel, H. P.
Asención Diez, Matías Damián
Wu, R.
Iglesias, Alberto Alvaro
Liu, D.
Ballicora, M. A.
author_role author
author2 Mascarenhas, R.
Patel, H. P.
Asención Diez, Matías Damián
Wu, R.
Iglesias, Alberto Alvaro
Liu, D.
Ballicora, M. A.
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv GLUCOSE-1-PHOSPHATE ADENYLYLTRANSFERASE
GLYCOGEN BIOSYNTHESIS
STARCH BIOSYNTHESIS
ALLOSTERISM
ENZYME EVOLUTION
GLUCAN BIOSYNTHESIS
topic GLUCOSE-1-PHOSPHATE ADENYLYLTRANSFERASE
GLYCOGEN BIOSYNTHESIS
STARCH BIOSYNTHESIS
ALLOSTERISM
ENZYME EVOLUTION
GLUCAN BIOSYNTHESIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The pathways for biosynthesis of glycogen inbacteria and starch in plants are evolutionarily andbiochemically related. They are regulated primarily by ADP?glucose pyrophosphorylase, which evolved to satisfy metabolic requirements of a particular organism. Despite the importance of these two pathways, little is known about the mechanism that controls pyrophosphorylase activity or the location of its allosteric sites. Here, we report pyruvate-bound crystal structures of ADP-glucose pyrophosphorylase from the bacterium Agrobacterium tumefaciens, identifying a previously elusive activator site for the enzyme. We found that the tetrameric enzyme binds two molecules of pyruvate in a planar conformation. Each binding site is located in a crevice between the C-terminal domains of two subunits where they stack via a distinct β-helix region. Pyruvate interacts with the side chain of Lys-43 and with the peptide backbone of Ser-328 and Gly-329 from both subunits. These structural insights led to the design of two variants with altered regulator properties. In one variant (K43A), the allosteric effect was absent, whereas in the other (G329D), the introduced Asp mimicked the presence of pyruvate. The latter generated an enzyme that was pre-activated and insensitive to further activation by pyruvate. Our study furnishes a deeper understanding of how glycogen biosynthesis is regulated in bacteria and the mechanism by which transgenic plants increased their starch production. These insights will facilitate rational approaches to enzyme engineering for starch production in crops of agricultural interest and will promote further study of allosteric signal transmission and molecular evolution in this important enzyme family.
Fil: Hill, B. L.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Mascarenhas, R.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Patel, H. P.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Asención Diez, Matías Damián. 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: Wu, R.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
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: Liu, D.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
Fil: Ballicora, M. A.. Dpt Of Chem And Biochemistry. Loyola University Chicago; Estados Unidos
description The pathways for biosynthesis of glycogen inbacteria and starch in plants are evolutionarily andbiochemically related. They are regulated primarily by ADP?glucose pyrophosphorylase, which evolved to satisfy metabolic requirements of a particular organism. Despite the importance of these two pathways, little is known about the mechanism that controls pyrophosphorylase activity or the location of its allosteric sites. Here, we report pyruvate-bound crystal structures of ADP-glucose pyrophosphorylase from the bacterium Agrobacterium tumefaciens, identifying a previously elusive activator site for the enzyme. We found that the tetrameric enzyme binds two molecules of pyruvate in a planar conformation. Each binding site is located in a crevice between the C-terminal domains of two subunits where they stack via a distinct β-helix region. Pyruvate interacts with the side chain of Lys-43 and with the peptide backbone of Ser-328 and Gly-329 from both subunits. These structural insights led to the design of two variants with altered regulator properties. In one variant (K43A), the allosteric effect was absent, whereas in the other (G329D), the introduced Asp mimicked the presence of pyruvate. The latter generated an enzyme that was pre-activated and insensitive to further activation by pyruvate. Our study furnishes a deeper understanding of how glycogen biosynthesis is regulated in bacteria and the mechanism by which transgenic plants increased their starch production. These insights will facilitate rational approaches to enzyme engineering for starch production in crops of agricultural interest and will promote further study of allosteric signal transmission and molecular evolution in this important enzyme family.
publishDate 2019
dc.date.none.fl_str_mv 2019-01
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/11336/118795
Hill, B. L.; Mascarenhas, R.; Patel, H. P.; Asención Diez, Matías Damián; Wu, R.; et al.; Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 294; 1-2019; 1338-1348
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/118795
identifier_str_mv Hill, B. L.; Mascarenhas, R.; Patel, H. P.; Asención Diez, Matías Damián; Wu, R.; et al.; Structural analysis reveals a pyruvate-binding activator site in the Agrobacterium tumefaciens ADP–glucose pyrophosphorylase; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 294; 1-2019; 1338-1348
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/lookup/doi/10.1074/jbc.RA118.004246
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.RA118.004246
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 12.982451

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