Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.

Autores
Duek, Paula D.; Wolosiuk, Ricardo Alejandro
Año de publicación
2001
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The stroma of higher plant chloroplasts contains two thioredoxins (Trx) with different specificity for the reduction of protein disulfide bonds. Based upon electrostatic features of domains that participate in the thiol/disulfide exchange, we prepared mutants of rapeseed Trx-m bearing opposite charges at a single position and subsequently analyzed their action on the activation of rapeseed chloroplast fructose 1,6-phosphate (CFBPase). The replacement of Pro-35 with lysine and glutamic residues enhanced and impaired, respectively, the stimulation of CFBPase relative to the wild-type and the P35A mutant. Furthermore, the shielding of electrostatic interactions with high concentrations of KCl greatly increased and concurrently made indistinguishable the affinity of all variants for CFBPase. The capacity to stimulate the enzyme activity likewise was enhanced concertedly by fructose-1,6-bisphosphate and Ca(2+) but, at variance with the action of KCl, remained sensitive to charges in the side chain of mutants. These results were consistent with a mechanism in which intermolecular electrostatic interactions and intramolecular non-covalent interactions control the formation of the non-covalent complex between reduced Trx and oxidized CFBPase and, in so doing, modulate the thiol/disulfide exchange.
Fil: Duek, Paula D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Wolosiuk, Ricardo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Materia
Chloroplast
Thioredoxin
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:11336/47750
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/47750
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.Duek, Paula D.Wolosiuk, Ricardo AlejandroChloroplastThioredoxinhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The stroma of higher plant chloroplasts contains two thioredoxins (Trx) with different specificity for the reduction of protein disulfide bonds. Based upon electrostatic features of domains that participate in the thiol/disulfide exchange, we prepared mutants of rapeseed Trx-m bearing opposite charges at a single position and subsequently analyzed their action on the activation of rapeseed chloroplast fructose 1,6-phosphate (CFBPase). The replacement of Pro-35 with lysine and glutamic residues enhanced and impaired, respectively, the stimulation of CFBPase relative to the wild-type and the P35A mutant. Furthermore, the shielding of electrostatic interactions with high concentrations of KCl greatly increased and concurrently made indistinguishable the affinity of all variants for CFBPase. The capacity to stimulate the enzyme activity likewise was enhanced concertedly by fructose-1,6-bisphosphate and Ca(2+) but, at variance with the action of KCl, remained sensitive to charges in the side chain of mutants. These results were consistent with a mechanism in which intermolecular electrostatic interactions and intramolecular non-covalent interactions control the formation of the non-covalent complex between reduced Trx and oxidized CFBPase and, in so doing, modulate the thiol/disulfide exchange.Fil: Duek, Paula D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Wolosiuk, Ricardo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaElsevier Science2001-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/47750Duek, Paula D.; Wolosiuk, Ricardo Alejandro; Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.; Elsevier Science; Biochimica Et Biophysica Acta-protein Structure And Molecular Enzymology; 1546; 2; 4-2001; 299-3110167-48381878-2434CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0167483801001509info:eu-repo/semantics/altIdentifier/doi/10.1016/S0167-4838(01)00150-9info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:48:13Zoai:ri.conicet.gov.ar:11336/47750instacron: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 11:48:14.172CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
title Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
spellingShingle Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
Duek, Paula D.
Chloroplast
Thioredoxin
title_short Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
title_full Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
title_fullStr Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
title_full_unstemmed Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
title_sort Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.
dc.creator.none.fl_str_mv Duek, Paula D.
Wolosiuk, Ricardo Alejandro
author Duek, Paula D.
author_facet Duek, Paula D.
Wolosiuk, Ricardo Alejandro
author_role author
author2 Wolosiuk, Ricardo Alejandro
author2_role author
dc.subject.none.fl_str_mv Chloroplast
Thioredoxin
topic Chloroplast
Thioredoxin
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 stroma of higher plant chloroplasts contains two thioredoxins (Trx) with different specificity for the reduction of protein disulfide bonds. Based upon electrostatic features of domains that participate in the thiol/disulfide exchange, we prepared mutants of rapeseed Trx-m bearing opposite charges at a single position and subsequently analyzed their action on the activation of rapeseed chloroplast fructose 1,6-phosphate (CFBPase). The replacement of Pro-35 with lysine and glutamic residues enhanced and impaired, respectively, the stimulation of CFBPase relative to the wild-type and the P35A mutant. Furthermore, the shielding of electrostatic interactions with high concentrations of KCl greatly increased and concurrently made indistinguishable the affinity of all variants for CFBPase. The capacity to stimulate the enzyme activity likewise was enhanced concertedly by fructose-1,6-bisphosphate and Ca(2+) but, at variance with the action of KCl, remained sensitive to charges in the side chain of mutants. These results were consistent with a mechanism in which intermolecular electrostatic interactions and intramolecular non-covalent interactions control the formation of the non-covalent complex between reduced Trx and oxidized CFBPase and, in so doing, modulate the thiol/disulfide exchange.
Fil: Duek, Paula D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Wolosiuk, Ricardo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
description The stroma of higher plant chloroplasts contains two thioredoxins (Trx) with different specificity for the reduction of protein disulfide bonds. Based upon electrostatic features of domains that participate in the thiol/disulfide exchange, we prepared mutants of rapeseed Trx-m bearing opposite charges at a single position and subsequently analyzed their action on the activation of rapeseed chloroplast fructose 1,6-phosphate (CFBPase). The replacement of Pro-35 with lysine and glutamic residues enhanced and impaired, respectively, the stimulation of CFBPase relative to the wild-type and the P35A mutant. Furthermore, the shielding of electrostatic interactions with high concentrations of KCl greatly increased and concurrently made indistinguishable the affinity of all variants for CFBPase. The capacity to stimulate the enzyme activity likewise was enhanced concertedly by fructose-1,6-bisphosphate and Ca(2+) but, at variance with the action of KCl, remained sensitive to charges in the side chain of mutants. These results were consistent with a mechanism in which intermolecular electrostatic interactions and intramolecular non-covalent interactions control the formation of the non-covalent complex between reduced Trx and oxidized CFBPase and, in so doing, modulate the thiol/disulfide exchange.
publishDate 2001
dc.date.none.fl_str_mv 2001-04
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/11336/47750
Duek, Paula D.; Wolosiuk, Ricardo Alejandro; Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.; Elsevier Science; Biochimica Et Biophysica Acta-protein Structure And Molecular Enzymology; 1546; 2; 4-2001; 299-311
0167-4838
1878-2434
CONICET Digital
CONICET
url http://hdl.handle.net/11336/47750
identifier_str_mv Duek, Paula D.; Wolosiuk, Ricardo Alejandro; Rapeseed chloroplast thioredoxin-m. Modulation of the affinity for target proteins.; Elsevier Science; Biochimica Et Biophysica Acta-protein Structure And Molecular Enzymology; 1546; 2; 4-2001; 299-311
0167-4838
1878-2434
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.sciencedirect.com/science/article/pii/S0167483801001509
info:eu-repo/semantics/altIdentifier/doi/10.1016/S0167-4838(01)00150-9
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 Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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_ 1846782188241027072
score 13.199325

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