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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/118795
Ver los metadatos del registro completo
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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 Técnicas2025-09-29T10:40:27Zoai: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:34982025-09-29 10:40:28.073CONICET 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 |
13.070432 |