Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA

Autores
Corbalan, Natalia Soledad; Runti, Giulia; Adler, Conrado; Covaceuszach, Sonia; Ford, Robert C.; Lamba, Doriano; Beis, Konstantinos; Scocchi, Marco; Vincent, Paula Andrea
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
SbmA protein has been proposed as a dimeric secondary transporter. The protein is involved in the transport of microcins B17 and J25, bleomycin, proline-rich antimicrobial peptides, antisense peptide phosphorodiamidate morpholino oligomers, and peptide nucleic acids into the Escherichia coli cytoplasm. The sbmA homologue is found in a variety of bacteria, though the physiological role of the protein is hitherto unknown. In this work, we carried out a functional and structural analysis to determine which amino acids are critical for the transport properties of SbmA. We created a set of 15 site-directed sbmA mutants in which single conserved amino acids were replaced by glycine residues. Our work demonstrated that strains carrying the site-directed mutants V102G, F219G, and E276G had a null phenotype for SbmA transport functions. In contrast, strains carrying the single point mutants W19G, W53G, F60G, S69G, N155G, R190, L233G, A344G, T255G, N308G, and R385G showed transport capacities indistinguishable from those of strains harboring a wild-type sbmA. The strain carrying the Y116G mutant exhibited mixed phenotypic characteristics. We also demonstrated that those sbmA mutants with severely impaired transport capacity showed a dominant negative phenotype. Electron microscopy data and in silico three-dimensional (3D) homology modeling support the idea that SbmA forms a homodimeric complex, closely resembling the membrane-spanning region of the ATP-binding cassette transporter family. Direct mapping of the sbmA single point mutants on the protein surface allowed us to explain the observed phenotypic differences in transport ability.
Fil: Corbalan, Natalia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina
Fil: Runti, Giulia. Università degli Studi di Trieste; Italia
Fil: Adler, Conrado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina
Fil: Covaceuszach, Sonia. Consiglio Nazionale delle Ricerche; Italia
Fil: Ford, Robert C.. University of Manchester; Reino Unido
Fil: Lamba, Doriano. Consiglio Nazionale delle Ricerche; Italia
Fil: Beis, Konstantinos. Imperial College London; Reino Unido
Fil: Scocchi, Marco. Università degli Studi di Trieste; Italia
Fil: Vincent, Paula Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina
Materia
Sbma
Function
Structure
Dimer
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:11336/22833
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/22833
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Functional and Structural Study of the Dimeric Inner Membrane Protein SbmACorbalan, Natalia SoledadRunti, GiuliaAdler, ConradoCovaceuszach, SoniaFord, Robert C.Lamba, DorianoBeis, KonstantinosScocchi, MarcoVincent, Paula AndreaSbmaFunctionStructureDimerhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1SbmA protein has been proposed as a dimeric secondary transporter. The protein is involved in the transport of microcins B17 and J25, bleomycin, proline-rich antimicrobial peptides, antisense peptide phosphorodiamidate morpholino oligomers, and peptide nucleic acids into the Escherichia coli cytoplasm. The sbmA homologue is found in a variety of bacteria, though the physiological role of the protein is hitherto unknown. In this work, we carried out a functional and structural analysis to determine which amino acids are critical for the transport properties of SbmA. We created a set of 15 site-directed sbmA mutants in which single conserved amino acids were replaced by glycine residues. Our work demonstrated that strains carrying the site-directed mutants V102G, F219G, and E276G had a null phenotype for SbmA transport functions. In contrast, strains carrying the single point mutants W19G, W53G, F60G, S69G, N155G, R190, L233G, A344G, T255G, N308G, and R385G showed transport capacities indistinguishable from those of strains harboring a wild-type sbmA. The strain carrying the Y116G mutant exhibited mixed phenotypic characteristics. We also demonstrated that those sbmA mutants with severely impaired transport capacity showed a dominant negative phenotype. Electron microscopy data and in silico three-dimensional (3D) homology modeling support the idea that SbmA forms a homodimeric complex, closely resembling the membrane-spanning region of the ATP-binding cassette transporter family. Direct mapping of the sbmA single point mutants on the protein surface allowed us to explain the observed phenotypic differences in transport ability.Fil: Corbalan, Natalia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; ArgentinaFil: Runti, Giulia. Università degli Studi di Trieste; ItaliaFil: Adler, Conrado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; ArgentinaFil: Covaceuszach, Sonia. Consiglio Nazionale delle Ricerche; ItaliaFil: Ford, Robert C.. University of Manchester; Reino UnidoFil: Lamba, Doriano. Consiglio Nazionale delle Ricerche; ItaliaFil: Beis, Konstantinos. Imperial College London; Reino UnidoFil: Scocchi, Marco. Università degli Studi di Trieste; ItaliaFil: Vincent, Paula Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; ArgentinaAmerican Society for Microbiology2013-09info: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/11336/22833Corbalan, Natalia Soledad; Runti, Giulia; Adler, Conrado; Covaceuszach, Sonia; Ford, Robert C.; et al.; Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA; American Society for Microbiology; Journal Of Bacteriology; 195; 23; 9-2013; 5352-53610021-91931098-5530CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1128/JB.00824-13info:eu-repo/semantics/altIdentifier/url/http://jb.asm.org/content/195/23/5352info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:58:30Zoai:ri.conicet.gov.ar:11336/22833instacron: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 09:58:31.266CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
title Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
spellingShingle Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
Corbalan, Natalia Soledad
Sbma
Function
Structure
Dimer
title_short Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
title_full Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
title_fullStr Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
title_full_unstemmed Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
title_sort Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA
dc.creator.none.fl_str_mv Corbalan, Natalia Soledad
Runti, Giulia
Adler, Conrado
Covaceuszach, Sonia
Ford, Robert C.
Lamba, Doriano
Beis, Konstantinos
Scocchi, Marco
Vincent, Paula Andrea
author Corbalan, Natalia Soledad
author_facet Corbalan, Natalia Soledad
Runti, Giulia
Adler, Conrado
Covaceuszach, Sonia
Ford, Robert C.
Lamba, Doriano
Beis, Konstantinos
Scocchi, Marco
Vincent, Paula Andrea
author_role author
author2 Runti, Giulia
Adler, Conrado
Covaceuszach, Sonia
Ford, Robert C.
Lamba, Doriano
Beis, Konstantinos
Scocchi, Marco
Vincent, Paula Andrea
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Sbma
Function
Structure
Dimer
topic Sbma
Function
Structure
Dimer
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv SbmA protein has been proposed as a dimeric secondary transporter. The protein is involved in the transport of microcins B17 and J25, bleomycin, proline-rich antimicrobial peptides, antisense peptide phosphorodiamidate morpholino oligomers, and peptide nucleic acids into the Escherichia coli cytoplasm. The sbmA homologue is found in a variety of bacteria, though the physiological role of the protein is hitherto unknown. In this work, we carried out a functional and structural analysis to determine which amino acids are critical for the transport properties of SbmA. We created a set of 15 site-directed sbmA mutants in which single conserved amino acids were replaced by glycine residues. Our work demonstrated that strains carrying the site-directed mutants V102G, F219G, and E276G had a null phenotype for SbmA transport functions. In contrast, strains carrying the single point mutants W19G, W53G, F60G, S69G, N155G, R190, L233G, A344G, T255G, N308G, and R385G showed transport capacities indistinguishable from those of strains harboring a wild-type sbmA. The strain carrying the Y116G mutant exhibited mixed phenotypic characteristics. We also demonstrated that those sbmA mutants with severely impaired transport capacity showed a dominant negative phenotype. Electron microscopy data and in silico three-dimensional (3D) homology modeling support the idea that SbmA forms a homodimeric complex, closely resembling the membrane-spanning region of the ATP-binding cassette transporter family. Direct mapping of the sbmA single point mutants on the protein surface allowed us to explain the observed phenotypic differences in transport ability.
Fil: Corbalan, Natalia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina
Fil: Runti, Giulia. Università degli Studi di Trieste; Italia
Fil: Adler, Conrado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina
Fil: Covaceuszach, Sonia. Consiglio Nazionale delle Ricerche; Italia
Fil: Ford, Robert C.. University of Manchester; Reino Unido
Fil: Lamba, Doriano. Consiglio Nazionale delle Ricerche; Italia
Fil: Beis, Konstantinos. Imperial College London; Reino Unido
Fil: Scocchi, Marco. Università degli Studi di Trieste; Italia
Fil: Vincent, Paula Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina
description SbmA protein has been proposed as a dimeric secondary transporter. The protein is involved in the transport of microcins B17 and J25, bleomycin, proline-rich antimicrobial peptides, antisense peptide phosphorodiamidate morpholino oligomers, and peptide nucleic acids into the Escherichia coli cytoplasm. The sbmA homologue is found in a variety of bacteria, though the physiological role of the protein is hitherto unknown. In this work, we carried out a functional and structural analysis to determine which amino acids are critical for the transport properties of SbmA. We created a set of 15 site-directed sbmA mutants in which single conserved amino acids were replaced by glycine residues. Our work demonstrated that strains carrying the site-directed mutants V102G, F219G, and E276G had a null phenotype for SbmA transport functions. In contrast, strains carrying the single point mutants W19G, W53G, F60G, S69G, N155G, R190, L233G, A344G, T255G, N308G, and R385G showed transport capacities indistinguishable from those of strains harboring a wild-type sbmA. The strain carrying the Y116G mutant exhibited mixed phenotypic characteristics. We also demonstrated that those sbmA mutants with severely impaired transport capacity showed a dominant negative phenotype. Electron microscopy data and in silico three-dimensional (3D) homology modeling support the idea that SbmA forms a homodimeric complex, closely resembling the membrane-spanning region of the ATP-binding cassette transporter family. Direct mapping of the sbmA single point mutants on the protein surface allowed us to explain the observed phenotypic differences in transport ability.
publishDate 2013
dc.date.none.fl_str_mv 2013-09
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/11336/22833
Corbalan, Natalia Soledad; Runti, Giulia; Adler, Conrado; Covaceuszach, Sonia; Ford, Robert C.; et al.; Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA; American Society for Microbiology; Journal Of Bacteriology; 195; 23; 9-2013; 5352-5361
0021-9193
1098-5530
CONICET Digital
CONICET
url http://hdl.handle.net/11336/22833
identifier_str_mv Corbalan, Natalia Soledad; Runti, Giulia; Adler, Conrado; Covaceuszach, Sonia; Ford, Robert C.; et al.; Functional and Structural Study of the Dimeric Inner Membrane Protein SbmA; American Society for Microbiology; Journal Of Bacteriology; 195; 23; 9-2013; 5352-5361
0021-9193
1098-5530
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1128/JB.00824-13
info:eu-repo/semantics/altIdentifier/url/http://jb.asm.org/content/195/23/5352
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 American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
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 13.070432

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