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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/51966
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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 Técnicas2025-09-29T09:58:01Zoai: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:34982025-09-29 09:58:02.033CONICET 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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1844613732146085888 |
score |
13.070432 |