Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells

Autores
Gomez-Mejiba, Sandra Esther; Zhai, Zili; Muñoz, Marcos David; Della Vedova, Maria Cecilia; Ranguelova, Kalina; Ashby, Michael T.; Ramirez, Dario
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A number of post-translational oxidative modifications of the enzyme “cell-redox sensor” glyceraldehyde-3- phosphate dehydrogenase (GAPDH) have been reported. These modifications affect GAPDH structure, function, and cell fate; however no free-radical mechanisms have been reported in these processes. Herein we used the nitrone 5,5-dimethyl-1-pyrroline N-oxide (DMPO)-based spin trapping techniques to examine a novel free radical mechanism that causes GAPDH inactivation and aggregation in RAW264.7 cells primed with lipopolysaccharide (LPS). In these primed cells, GAPDH is oxidized by myeloperoxidase (MPO)-derived hypochlorous acid (HOCl) resulting in loss of enzyme activity and aggregation, accumulation of lactate and cell death. Due to the close spatial and physical proximity between MPO and GAPDH, and the oxidizing potential of HOCl, it may be the main species that triggers radicalization of GAPDH that ultimately results in enzyme aggregation and inactivation in LPS-primed macrophages. Lysine residues are the primary radicalization sites formed upon reaction of HOCl with the enzyme. Our data highlight the important relationship between radicalization of GAPDH and fate of stressed cells, which might help teasing out the cell response to stress at sites of inflammation.
Fil: Gomez-Mejiba, Sandra Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina
Fil: Zhai, Zili. University of Colorado; Estados Unidos
Fil: Muñoz, Marcos David. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina
Fil: Della Vedova, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina
Fil: Ranguelova, Kalina. Bruker BioSpin Corporation; Estados Unidos
Fil: Ashby, Michael T.. University of Oklahoma; Estados Unidos
Fil: Ramirez, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina
Materia
GAPDH
MPO
HOCL
Oxidative modification
Macrophage
Lipopolysaccharide
Reactive chemical species
Cell death
Protein radical
Immuno-spin trapping
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:11336/51966
Acceder
id CONICETDig_54327ef1e66d3e7fd5485687146b14e5
oai_identifier_str oai:ri.conicet.gov.ar:11336/51966
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living CellsGomez-Mejiba, Sandra EstherZhai, ZiliMuñoz, Marcos DavidDella Vedova, Maria CeciliaRanguelova, KalinaAshby, Michael T.Ramirez, DarioGAPDHMPOHOCLOxidative modificationMacrophageLipopolysaccharideReactive chemical speciesCell deathProtein radicalImmuno-spin trappinghttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1A number of post-translational oxidative modifications of the enzyme “cell-redox sensor” glyceraldehyde-3- phosphate dehydrogenase (GAPDH) have been reported. These modifications affect GAPDH structure, function, and cell fate; however no free-radical mechanisms have been reported in these processes. Herein we used the nitrone 5,5-dimethyl-1-pyrroline N-oxide (DMPO)-based spin trapping techniques to examine a novel free radical mechanism that causes GAPDH inactivation and aggregation in RAW264.7 cells primed with lipopolysaccharide (LPS). In these primed cells, GAPDH is oxidized by myeloperoxidase (MPO)-derived hypochlorous acid (HOCl) resulting in loss of enzyme activity and aggregation, accumulation of lactate and cell death. Due to the close spatial and physical proximity between MPO and GAPDH, and the oxidizing potential of HOCl, it may be the main species that triggers radicalization of GAPDH that ultimately results in enzyme aggregation and inactivation in LPS-primed macrophages. Lysine residues are the primary radicalization sites formed upon reaction of HOCl with the enzyme. Our data highlight the important relationship between radicalization of GAPDH and fate of stressed cells, which might help teasing out the cell response to stress at sites of inflammation.Fil: Gomez-Mejiba, Sandra Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; ArgentinaFil: Zhai, Zili. University of Colorado; Estados UnidosFil: Muñoz, Marcos David. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; ArgentinaFil: Della Vedova, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; ArgentinaFil: Ranguelova, Kalina. Bruker BioSpin Corporation; Estados UnidosFil: Ashby, Michael T.. University of Oklahoma; Estados UnidosFil: Ramirez, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; ArgentinaOMICS International2015-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/51966Gomez-Mejiba, Sandra Esther; Zhai, Zili; Muñoz, Marcos David; Della Vedova, Maria Cecilia; Ranguelova, Kalina; et al.; Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells; OMICS International; Enzyme Engineering; 4; 2; 12-2015; 134-1392329-6674CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.omicsgroup.org/journals/radicalization-of-glyceraldehyde3phosphate-dehydrogenase-by-hocl-inliving-cells-2329-6674-1000134.php?aid=64284info:eu-repo/semantics/altIdentifier/doi/10.4172/2329-6674.1000134info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:53:26Zoai:ri.conicet.gov.ar:11336/51966instacron: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:34982025-10-15 14:53:26.93CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
title Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
spellingShingle Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
Gomez-Mejiba, Sandra Esther
GAPDH
MPO
HOCL
Oxidative modification
Macrophage
Lipopolysaccharide
Reactive chemical species
Cell death
Protein radical
Immuno-spin trapping
title_short Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
title_full Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
title_fullStr Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
title_full_unstemmed Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
title_sort Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells
dc.creator.none.fl_str_mv Gomez-Mejiba, Sandra Esther
Zhai, Zili
Muñoz, Marcos David
Della Vedova, Maria Cecilia
Ranguelova, Kalina
Ashby, Michael T.
Ramirez, Dario
author Gomez-Mejiba, Sandra Esther
author_facet Gomez-Mejiba, Sandra Esther
Zhai, Zili
Muñoz, Marcos David
Della Vedova, Maria Cecilia
Ranguelova, Kalina
Ashby, Michael T.
Ramirez, Dario
author_role author
author2 Zhai, Zili
Muñoz, Marcos David
Della Vedova, Maria Cecilia
Ranguelova, Kalina
Ashby, Michael T.
Ramirez, Dario
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv GAPDH
MPO
HOCL
Oxidative modification
Macrophage
Lipopolysaccharide
Reactive chemical species
Cell death
Protein radical
Immuno-spin trapping
topic GAPDH
MPO
HOCL
Oxidative modification
Macrophage
Lipopolysaccharide
Reactive chemical species
Cell death
Protein radical
Immuno-spin trapping
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv A number of post-translational oxidative modifications of the enzyme “cell-redox sensor” glyceraldehyde-3- phosphate dehydrogenase (GAPDH) have been reported. These modifications affect GAPDH structure, function, and cell fate; however no free-radical mechanisms have been reported in these processes. Herein we used the nitrone 5,5-dimethyl-1-pyrroline N-oxide (DMPO)-based spin trapping techniques to examine a novel free radical mechanism that causes GAPDH inactivation and aggregation in RAW264.7 cells primed with lipopolysaccharide (LPS). In these primed cells, GAPDH is oxidized by myeloperoxidase (MPO)-derived hypochlorous acid (HOCl) resulting in loss of enzyme activity and aggregation, accumulation of lactate and cell death. Due to the close spatial and physical proximity between MPO and GAPDH, and the oxidizing potential of HOCl, it may be the main species that triggers radicalization of GAPDH that ultimately results in enzyme aggregation and inactivation in LPS-primed macrophages. Lysine residues are the primary radicalization sites formed upon reaction of HOCl with the enzyme. Our data highlight the important relationship between radicalization of GAPDH and fate of stressed cells, which might help teasing out the cell response to stress at sites of inflammation.
Fil: Gomez-Mejiba, Sandra Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina
Fil: Zhai, Zili. University of Colorado; Estados Unidos
Fil: Muñoz, Marcos David. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina
Fil: Della Vedova, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Ciencias de la Salud; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina
Fil: Ranguelova, Kalina. Bruker BioSpin Corporation; Estados Unidos
Fil: Ashby, Michael T.. University of Oklahoma; Estados Unidos
Fil: Ramirez, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina
description A number of post-translational oxidative modifications of the enzyme “cell-redox sensor” glyceraldehyde-3- phosphate dehydrogenase (GAPDH) have been reported. These modifications affect GAPDH structure, function, and cell fate; however no free-radical mechanisms have been reported in these processes. Herein we used the nitrone 5,5-dimethyl-1-pyrroline N-oxide (DMPO)-based spin trapping techniques to examine a novel free radical mechanism that causes GAPDH inactivation and aggregation in RAW264.7 cells primed with lipopolysaccharide (LPS). In these primed cells, GAPDH is oxidized by myeloperoxidase (MPO)-derived hypochlorous acid (HOCl) resulting in loss of enzyme activity and aggregation, accumulation of lactate and cell death. Due to the close spatial and physical proximity between MPO and GAPDH, and the oxidizing potential of HOCl, it may be the main species that triggers radicalization of GAPDH that ultimately results in enzyme aggregation and inactivation in LPS-primed macrophages. Lysine residues are the primary radicalization sites formed upon reaction of HOCl with the enzyme. Our data highlight the important relationship between radicalization of GAPDH and fate of stressed cells, which might help teasing out the cell response to stress at sites of inflammation.
publishDate 2015
dc.date.none.fl_str_mv 2015-12
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/11336/51966
Gomez-Mejiba, Sandra Esther; Zhai, Zili; Muñoz, Marcos David; Della Vedova, Maria Cecilia; Ranguelova, Kalina; et al.; Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells; OMICS International; Enzyme Engineering; 4; 2; 12-2015; 134-139
2329-6674
CONICET Digital
CONICET
url http://hdl.handle.net/11336/51966
identifier_str_mv Gomez-Mejiba, Sandra Esther; Zhai, Zili; Muñoz, Marcos David; Della Vedova, Maria Cecilia; Ranguelova, Kalina; et al.; Radicalization of Glyceraldehyde-3-Phosphate Dehydrogenase by HOCl in Living Cells; OMICS International; Enzyme Engineering; 4; 2; 12-2015; 134-139
2329-6674
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.omicsgroup.org/journals/radicalization-of-glyceraldehyde3phosphate-dehydrogenase-by-hocl-inliving-cells-2329-6674-1000134.php?aid=64284
info:eu-repo/semantics/altIdentifier/doi/10.4172/2329-6674.1000134
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
application/pdf
dc.publisher.none.fl_str_mv OMICS International
publisher.none.fl_str_mv OMICS International
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 13.22299

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