Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119

An NADPH-glutathione reductase (EC 1.6.4.2) has been purified 6,000-fold to electrophoretic homogeneity from the filamentous cyanobacterium Anabaena sp. strain 7119. The purified enzyme exhibits a specific activity of 249 U/mg and is characterized by being a dimeric flavin adenine dinucleotide-conta...

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Autores: Losada Villasante, Manuel, Serrano Delgado, Aurelio, Rivas Florido, Joaquín
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:1984
País:España
Institución:Universidad de Sevilla (US)
Repositorio:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/26063
Acceso en línea:http://hdl.handle.net/11441/26063
Access Level:acceso abierto
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spelling Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119Losada Villasante, ManuelSerrano Delgado, AurelioRivas Florido, JoaquínAn NADPH-glutathione reductase (EC 1.6.4.2) has been purified 6,000-fold to electrophoretic homogeneity from the filamentous cyanobacterium Anabaena sp. strain 7119. The purified enzyme exhibits a specific activity of 249 U/mg and is characterized by being a dimeric flavin adenine dinucleotide-containing protein with a ratio of absorbance at 280 nm to absorbance at 462 nm of 5.8, a native molecular weight of 104,000, a Stokes radius of 4.13 nm, and a pI of 4.02. The enzyme activity is inhibited by sulfhydryl reagents and heavy-metal ions, especially in the presence of NADPH, with oxidized glutathione behaving as a protective agent. As is the case with the same enzyme from other sources, the kinetic data are consistent with a branched mechanism. Nevertheless, the cyanobacterial enzyme presents three distinctive features with respect to that isolated from non-photosynthetic organisms: (i) absolute specificity for NADPH, (ii) an alkaline optimum pH value of ca. 9.0, and (iii) strong acidic character of the protein, as estimated by column chromatofocusing. The kinetic parameters are very similar to those found for the chloroplast enzyme, but the molecular weight is lower, being comparable to that of non-photosynthetic microorganisms. A protective function, analogous to that assigned to the chloroplast enzyme, is suggested.American Society for MicrobiologyBioquímica Vegetal y Biología Molecular1984info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/11441/26063reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésJournal of Bacteriology, 158 (1), 317-324.http://jb.asm.org/content/158/1/317.full.pdf+htmlinfo:eu-repo/semantics/openAccessoai:idus.us.es:11441/260632026-06-17T12:51:07Z
dc.title.none.fl_str_mv Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
title Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
spellingShingle Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
Losada Villasante, Manuel
title_short Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
title_full Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
title_fullStr Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
title_full_unstemmed Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
title_sort Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119
dc.creator.none.fl_str_mv Losada Villasante, Manuel
Serrano Delgado, Aurelio
Rivas Florido, Joaquín
author Losada Villasante, Manuel
author_facet Losada Villasante, Manuel
Serrano Delgado, Aurelio
Rivas Florido, Joaquín
author_role author
author2 Serrano Delgado, Aurelio
Rivas Florido, Joaquín
author2_role author
author
dc.contributor.none.fl_str_mv Bioquímica Vegetal y Biología Molecular
description An NADPH-glutathione reductase (EC 1.6.4.2) has been purified 6,000-fold to electrophoretic homogeneity from the filamentous cyanobacterium Anabaena sp. strain 7119. The purified enzyme exhibits a specific activity of 249 U/mg and is characterized by being a dimeric flavin adenine dinucleotide-containing protein with a ratio of absorbance at 280 nm to absorbance at 462 nm of 5.8, a native molecular weight of 104,000, a Stokes radius of 4.13 nm, and a pI of 4.02. The enzyme activity is inhibited by sulfhydryl reagents and heavy-metal ions, especially in the presence of NADPH, with oxidized glutathione behaving as a protective agent. As is the case with the same enzyme from other sources, the kinetic data are consistent with a branched mechanism. Nevertheless, the cyanobacterial enzyme presents three distinctive features with respect to that isolated from non-photosynthetic organisms: (i) absolute specificity for NADPH, (ii) an alkaline optimum pH value of ca. 9.0, and (iii) strong acidic character of the protein, as estimated by column chromatofocusing. The kinetic parameters are very similar to those found for the chloroplast enzyme, but the molecular weight is lower, being comparable to that of non-photosynthetic microorganisms. A protective function, analogous to that assigned to the chloroplast enzyme, is suggested.
publishDate 1984
dc.date.none.fl_str_mv 1984
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11441/26063
url http://hdl.handle.net/11441/26063
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Journal of Bacteriology, 158 (1), 317-324.
http://jb.asm.org/content/158/1/317.full.pdf+html
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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