Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves

Autores
Gomez Casati, Diego Fabian; Sesma, Juliana; Iglesias, Alberto Alvaro
Año de publicación
2000
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
NADP-dependent, non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.9) from celery leaves was purified over 1200-fold to a specific activity of 35 units/mg protein, and its kinetic, regulatory and structural properties were characterized. The purified enzyme exhibited a homotetrameric structure with a subunit molecular mass of 54 kDa. A high specificity of the enzyme for the substrates NADP+ (K(m) = 7 μM) and D- glyceraldehyde-3-phosphate (K(m) = 127 μM) was observed. Maximal activity was determined at pH 8.5. The purified enzyme was highly unstable, requiring the addition of NADP+ or conditions of high ionic strength in the medium. A hysteretic behavior, with a lag phase of minutes, was observed during activity measurement of the enzyme preincubated in the absence of substrates. The lag was inversely proportional to the protein concentration during preincubation. The hysteretic parameters were affected by the substrates, KCl and mannitol among other compounds. Distinctively, incubation with NADP+ produced a near twofold activation of the enzyme. Results suggest that in alditol producing plants the enzyme plays a key role in the synthesis and partitioning of photoassimilates.
Fil: Gomez Casati, Diego Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Sesma, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; 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
Materia
ALDITOLS BIOSYNTHESIS
CARBON PARTITIONING
CELERY LEAVES
GLYCERALDEHYDE- 3-PHOSPHATE DEHYDROGENASE
NON-PHOSPHORYLATING
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:11336/85665
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/85665
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leavesGomez Casati, Diego FabianSesma, JulianaIglesias, Alberto AlvaroALDITOLS BIOSYNTHESISCARBON PARTITIONINGCELERY LEAVESGLYCERALDEHYDE- 3-PHOSPHATE DEHYDROGENASENON-PHOSPHORYLATINGhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1NADP-dependent, non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.9) from celery leaves was purified over 1200-fold to a specific activity of 35 units/mg protein, and its kinetic, regulatory and structural properties were characterized. The purified enzyme exhibited a homotetrameric structure with a subunit molecular mass of 54 kDa. A high specificity of the enzyme for the substrates NADP+ (K(m) = 7 μM) and D- glyceraldehyde-3-phosphate (K(m) = 127 μM) was observed. Maximal activity was determined at pH 8.5. The purified enzyme was highly unstable, requiring the addition of NADP+ or conditions of high ionic strength in the medium. A hysteretic behavior, with a lag phase of minutes, was observed during activity measurement of the enzyme preincubated in the absence of substrates. The lag was inversely proportional to the protein concentration during preincubation. The hysteretic parameters were affected by the substrates, KCl and mannitol among other compounds. Distinctively, incubation with NADP+ produced a near twofold activation of the enzyme. Results suggest that in alditol producing plants the enzyme plays a key role in the synthesis and partitioning of photoassimilates.Fil: Gomez Casati, Diego Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Sesma, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; 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; ArgentinaElsevier Ireland2000-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/85665Gomez Casati, Diego Fabian; Sesma, Juliana; Iglesias, Alberto Alvaro; Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves; Elsevier Ireland; Plant Science; 154; 2; 5-2000; 107-1150168-9452CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.ncbi.nlm.nih.gov/pubmed/?term=Structural+and+kinetic+characterization+of+NADP-dependent%2C+non-phosphorylating+glycerladehyde-3-phosphate+dehydrogenase+from+celery+leavesinfo:eu-repo/semantics/altIdentifier/doi/10.1016/S0168-9452(99)00241-1info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:15:32Zoai:ri.conicet.gov.ar:11336/85665instacron: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:34982025-09-10 13:15:32.889CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
title Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
spellingShingle Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
Gomez Casati, Diego Fabian
ALDITOLS BIOSYNTHESIS
CARBON PARTITIONING
CELERY LEAVES
GLYCERALDEHYDE- 3-PHOSPHATE DEHYDROGENASE
NON-PHOSPHORYLATING
title_short Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
title_full Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
title_fullStr Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
title_full_unstemmed Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
title_sort Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves
dc.creator.none.fl_str_mv Gomez Casati, Diego Fabian
Sesma, Juliana
Iglesias, Alberto Alvaro
author Gomez Casati, Diego Fabian
author_facet Gomez Casati, Diego Fabian
Sesma, Juliana
Iglesias, Alberto Alvaro
author_role author
author2 Sesma, Juliana
Iglesias, Alberto Alvaro
author2_role author
author
dc.subject.none.fl_str_mv ALDITOLS BIOSYNTHESIS
CARBON PARTITIONING
CELERY LEAVES
GLYCERALDEHYDE- 3-PHOSPHATE DEHYDROGENASE
NON-PHOSPHORYLATING
topic ALDITOLS BIOSYNTHESIS
CARBON PARTITIONING
CELERY LEAVES
GLYCERALDEHYDE- 3-PHOSPHATE DEHYDROGENASE
NON-PHOSPHORYLATING
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv NADP-dependent, non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.9) from celery leaves was purified over 1200-fold to a specific activity of 35 units/mg protein, and its kinetic, regulatory and structural properties were characterized. The purified enzyme exhibited a homotetrameric structure with a subunit molecular mass of 54 kDa. A high specificity of the enzyme for the substrates NADP+ (K(m) = 7 μM) and D- glyceraldehyde-3-phosphate (K(m) = 127 μM) was observed. Maximal activity was determined at pH 8.5. The purified enzyme was highly unstable, requiring the addition of NADP+ or conditions of high ionic strength in the medium. A hysteretic behavior, with a lag phase of minutes, was observed during activity measurement of the enzyme preincubated in the absence of substrates. The lag was inversely proportional to the protein concentration during preincubation. The hysteretic parameters were affected by the substrates, KCl and mannitol among other compounds. Distinctively, incubation with NADP+ produced a near twofold activation of the enzyme. Results suggest that in alditol producing plants the enzyme plays a key role in the synthesis and partitioning of photoassimilates.
Fil: Gomez Casati, Diego Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Sesma, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; 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
description NADP-dependent, non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.9) from celery leaves was purified over 1200-fold to a specific activity of 35 units/mg protein, and its kinetic, regulatory and structural properties were characterized. The purified enzyme exhibited a homotetrameric structure with a subunit molecular mass of 54 kDa. A high specificity of the enzyme for the substrates NADP+ (K(m) = 7 μM) and D- glyceraldehyde-3-phosphate (K(m) = 127 μM) was observed. Maximal activity was determined at pH 8.5. The purified enzyme was highly unstable, requiring the addition of NADP+ or conditions of high ionic strength in the medium. A hysteretic behavior, with a lag phase of minutes, was observed during activity measurement of the enzyme preincubated in the absence of substrates. The lag was inversely proportional to the protein concentration during preincubation. The hysteretic parameters were affected by the substrates, KCl and mannitol among other compounds. Distinctively, incubation with NADP+ produced a near twofold activation of the enzyme. Results suggest that in alditol producing plants the enzyme plays a key role in the synthesis and partitioning of photoassimilates.
publishDate 2000
dc.date.none.fl_str_mv 2000-05
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/11336/85665
Gomez Casati, Diego Fabian; Sesma, Juliana; Iglesias, Alberto Alvaro; Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves; Elsevier Ireland; Plant Science; 154; 2; 5-2000; 107-115
0168-9452
CONICET Digital
CONICET
url http://hdl.handle.net/11336/85665
identifier_str_mv Gomez Casati, Diego Fabian; Sesma, Juliana; Iglesias, Alberto Alvaro; Structural and kinetic characterization of NADP-dependent, non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves; Elsevier Ireland; Plant Science; 154; 2; 5-2000; 107-115
0168-9452
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.ncbi.nlm.nih.gov/pubmed/?term=Structural+and+kinetic+characterization+of+NADP-dependent%2C+non-phosphorylating+glycerladehyde-3-phosphate+dehydrogenase+from+celery+leaves
info:eu-repo/semantics/altIdentifier/doi/10.1016/S0168-9452(99)00241-1
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
application/pdf
dc.publisher.none.fl_str_mv Elsevier Ireland
publisher.none.fl_str_mv Elsevier Ireland
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
_version_ 1842980838703104000
score 12.993085

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