Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli

Detalhes bibliográficos
Autor(a) principal: Alvarez, Nelson Alexander Araujo
Data de Publicação: 2024
Outros Autores: Veloso, Marcelo, Pouchucq, Luis
Tipo de documento: preprint
Idioma: spa
Título da fonte: SciELO Preprints
Texto Completo: https://preprints.scielo.org/index.php/scielo/preprint/view/7828
Resumo: FtsZ is a bacterial divisome protein responsible for Z-ring formation in cytokinesis. Characterization of Escherichia coli FtsZ protein oligomers (EcFtsZ) in native conditions is defiance because the protein is found as a multi-oligomer in self-association-dissociation equilibrium. We characterize the trimeric state of EcFtsZ through native PAGE, gel filtration chromatography and sucrose gradient techniques combined with chemical cross-linking. The filtration results indicate that the EcFtsZ trimer has a molecular mass of 131 kDa and a filtration friction coefficient (Rs/Rmin) equal to 1.9, while the theoretical filtration friction coefficient (fn/f1) calculated for a linear trimer yielded a value equal to 1.8 very close to the experimental value. On the other side, formaldehyde-crosslinked EcFtsZ showed a band 128 kDa recognized by anti-FtsZ antibodies, and a sedimentation friction coefficient (Smax/S20,w) equal to 1.9, while the theoretical sedimentation friction coefficient (Sn/S1) calculated for a triangular trimer was equal to the experimental value. These results suggest that EcFtsZ has two homotrimeric structures (linear and triangular). Finally, we report the aggregation of EcFtsZ at micromolar concentrations of Capsaicin without GTP and Mg2+.
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spelling Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coliCARACTERIZACIÓN BIOQUÍMICA DE DOS TRÍMEROS DE LA PROTEÍNA DE LA DIVISIÓN CELULAR BACTERIANA (FtsZ) DE E. coli.FtsZ ProteinTrimersNative PAGEFriction coefficientCapsaicinProteína FtsZTrímerosPAGE nativoCapsaicinaCoeficiente de fricciónFtsZ is a bacterial divisome protein responsible for Z-ring formation in cytokinesis. Characterization of Escherichia coli FtsZ protein oligomers (EcFtsZ) in native conditions is defiance because the protein is found as a multi-oligomer in self-association-dissociation equilibrium. We characterize the trimeric state of EcFtsZ through native PAGE, gel filtration chromatography and sucrose gradient techniques combined with chemical cross-linking. The filtration results indicate that the EcFtsZ trimer has a molecular mass of 131 kDa and a filtration friction coefficient (Rs/Rmin) equal to 1.9, while the theoretical filtration friction coefficient (fn/f1) calculated for a linear trimer yielded a value equal to 1.8 very close to the experimental value. On the other side, formaldehyde-crosslinked EcFtsZ showed a band 128 kDa recognized by anti-FtsZ antibodies, and a sedimentation friction coefficient (Smax/S20,w) equal to 1.9, while the theoretical sedimentation friction coefficient (Sn/S1) calculated for a triangular trimer was equal to the experimental value. These results suggest that EcFtsZ has two homotrimeric structures (linear and triangular). Finally, we report the aggregation of EcFtsZ at micromolar concentrations of Capsaicin without GTP and Mg2+.La FtsZ es una proteína del divisoma bacteriano responsable de la formación del anillo Z en la citocinesis. La caracterización de los oligómeros de la proteína FtsZ de Escherichia coli (EcFtsZ) en su estado nativo es un desafío porque la proteína se encuentra como un multioligómero en equilibrio de autoasociación-disociación. Nosotros caracterizamos el estado trimérico de la EcFtsZ a través de técnicas de PAGE nativo, cromatografía de filtración en gel y gradiente de sacarosa combinada con entrecruzamiento químico. Los resultados por filtración indican que el trímero de EcFtsZ tiene una masa molecular de 131 kDa y un coeficiente de fricción por filtración (Rs/Rmin) igual a 1.9, mientras que el coeficiente de fricción por filtración teórico (fn/f1) calculado para un trímero lineal arrojó un valor igual a 1.8 que está muy cercano al valor experimental. Por otro lado, la EcFtsZ entrecruzada con formaldehído presentó una banda de 128 kDa reconocida por anticuerpos anti-FtsZ y un coeficiente de fricción por sedimentación (Smax/S20,w) igual a 1.9, mientras que el coeficiente de fricción por sedimentación teórico (Sn/S1) calculado para un trímero triangular resultó igual a valor experimental. Estos resultados sugieren que la EcFtsZ tiene dos estructuras homotriméricas (lineal y triangular). Finalmente, reportamos la agregación de la EcFtsZ a concentraciones micromolares de Capsaicina sin GTP y Mg2+.SciELO PreprintsSciELO PreprintsSciELO Preprints2024-01-16info:eu-repo/semantics/preprintinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://preprints.scielo.org/index.php/scielo/preprint/view/782810.1590/SciELOPreprints.7828spahttps://preprints.scielo.org/index.php/scielo/article/view/7828/14759Copyright (c) 2024 Nelson Alexander Araujo Alvarez, Marcelo Veloso, Luis Pouchucqhttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessAlvarez, Nelson Alexander AraujoVeloso, MarceloPouchucq, Luisreponame:SciELO Preprintsinstname:Scientific Electronic Library Online (SCIELO)instacron:SCI2024-01-03T00:19:40Zoai:ops.preprints.scielo.org:preprint/7828Servidor de preprintshttps://preprints.scielo.org/index.php/scieloONGhttps://preprints.scielo.org/index.php/scielo/oaiscielo.submission@scielo.orgopendoar:2024-01-03T00:19:40SciELO Preprints - Scientific Electronic Library Online (SCIELO)false
dc.title.none.fl_str_mv Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
CARACTERIZACIÓN BIOQUÍMICA DE DOS TRÍMEROS DE LA PROTEÍNA DE LA DIVISIÓN CELULAR BACTERIANA (FtsZ) DE E. coli.
title Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
spellingShingle Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
Alvarez, Nelson Alexander Araujo
FtsZ Protein
Trimers
Native PAGE
Friction coefficient
Capsaicin
Proteína FtsZ
Trímeros
PAGE nativo
Capsaicina
Coeficiente de fricción
title_short Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
title_full Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
title_fullStr Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
title_full_unstemmed Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
title_sort Biochemical characterization of two trimers of the bacterial cell division protein (FtsZ) from E. coli
author Alvarez, Nelson Alexander Araujo
author_facet Alvarez, Nelson Alexander Araujo
Veloso, Marcelo
Pouchucq, Luis
author_role author
author2 Veloso, Marcelo
Pouchucq, Luis
author2_role author
author
dc.contributor.author.fl_str_mv Alvarez, Nelson Alexander Araujo
Veloso, Marcelo
Pouchucq, Luis
dc.subject.por.fl_str_mv FtsZ Protein
Trimers
Native PAGE
Friction coefficient
Capsaicin
Proteína FtsZ
Trímeros
PAGE nativo
Capsaicina
Coeficiente de fricción
topic FtsZ Protein
Trimers
Native PAGE
Friction coefficient
Capsaicin
Proteína FtsZ
Trímeros
PAGE nativo
Capsaicina
Coeficiente de fricción
description FtsZ is a bacterial divisome protein responsible for Z-ring formation in cytokinesis. Characterization of Escherichia coli FtsZ protein oligomers (EcFtsZ) in native conditions is defiance because the protein is found as a multi-oligomer in self-association-dissociation equilibrium. We characterize the trimeric state of EcFtsZ through native PAGE, gel filtration chromatography and sucrose gradient techniques combined with chemical cross-linking. The filtration results indicate that the EcFtsZ trimer has a molecular mass of 131 kDa and a filtration friction coefficient (Rs/Rmin) equal to 1.9, while the theoretical filtration friction coefficient (fn/f1) calculated for a linear trimer yielded a value equal to 1.8 very close to the experimental value. On the other side, formaldehyde-crosslinked EcFtsZ showed a band 128 kDa recognized by anti-FtsZ antibodies, and a sedimentation friction coefficient (Smax/S20,w) equal to 1.9, while the theoretical sedimentation friction coefficient (Sn/S1) calculated for a triangular trimer was equal to the experimental value. These results suggest that EcFtsZ has two homotrimeric structures (linear and triangular). Finally, we report the aggregation of EcFtsZ at micromolar concentrations of Capsaicin without GTP and Mg2+.
publishDate 2024
dc.date.none.fl_str_mv 2024-01-16
dc.type.driver.fl_str_mv info:eu-repo/semantics/preprint
info:eu-repo/semantics/publishedVersion
format preprint
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://preprints.scielo.org/index.php/scielo/preprint/view/7828
10.1590/SciELOPreprints.7828
url https://preprints.scielo.org/index.php/scielo/preprint/view/7828
identifier_str_mv 10.1590/SciELOPreprints.7828
dc.language.iso.fl_str_mv spa
language spa
dc.relation.none.fl_str_mv https://preprints.scielo.org/index.php/scielo/article/view/7828/14759
dc.rights.driver.fl_str_mv Copyright (c) 2024 Nelson Alexander Araujo Alvarez, Marcelo Veloso, Luis Pouchucq
https://creativecommons.org/licenses/by/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Copyright (c) 2024 Nelson Alexander Araujo Alvarez, Marcelo Veloso, Luis Pouchucq
https://creativecommons.org/licenses/by/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv SciELO Preprints
SciELO Preprints
SciELO Preprints
publisher.none.fl_str_mv SciELO Preprints
SciELO Preprints
SciELO Preprints
dc.source.none.fl_str_mv reponame:SciELO Preprints
instname:Scientific Electronic Library Online (SCIELO)
instacron:SCI
instname_str Scientific Electronic Library Online (SCIELO)
instacron_str SCI
institution SCI
reponame_str SciELO Preprints
collection SciELO Preprints
repository.name.fl_str_mv SciELO Preprints - Scientific Electronic Library Online (SCIELO)
repository.mail.fl_str_mv scielo.submission@scielo.org
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