Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues

Autores
Gonzalez, Lisandro Javier; Moreno, Diego Martin; Bonomo, Robert A.; Vila, Alejandro Jose
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Pseudomonas aeruginosa is one of the most virulent and resistant non-fermenting Gram-negative pathogens in the clinic. Unfortunately, P. aeruginosa has acquired genes encoding metallo-beta-lactamases (MBLs), enzymes able to hydrolyze most beta-lactam antibiotics. SPM-1 is an MBL produced only by P. aeruginosa, while other MBLs are found in different bacteria. Despite similar active sites, the resistance profile of MBLs towards beta-lactams changes from one enzyme to the other. SPM-1 is unique among pathogen-associated MBLs in that in that it contains "atypical" second sphere residues (S84, G121). Codon randomization on these positions and further selection of resistance-conferring mutants was performed. MICs, periplasmic enzymatic activity, Zn(II) requirements, and protein stability was assessed. Our results indicated that identity of second sphere residues modulates the substrate preferences and the resistance profile of SPM-1 expressed in P. aeruginosa. The second sphere residues found in wild type SPM-1 give rise to a substrate selectivity that is observed only in the periplasmic environment. These residues also allow SPM-1 to confer resistance in P. aeruginosa under Zn(II)-limiting conditions, such as those expected under infection. By optimizing the catalytic efficiency towards beta-lactam antibiotics, the enzyme stability and the Zn(II) binding features, molecular evolution meets the specific needs of a pathogenic bacterial host by means of substitutions outside the active site.
Fil: Gonzalez, Lisandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; Argentina
Fil: Bonomo, Robert A.. Case Western Reserve University; Estados Unidos
Fil: Vila, Alejandro Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Materia
Antibiotics
Bacterial Pathogens
Codons
Hydrogen bonding
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/7696

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network_name_str CONICET Digital (CONICET)
spelling Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere ResiduesGonzalez, Lisandro JavierMoreno, Diego MartinBonomo, Robert A.Vila, Alejandro JoseAntibioticsBacterial PathogensCodonsHydrogen bondinghttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Pseudomonas aeruginosa is one of the most virulent and resistant non-fermenting Gram-negative pathogens in the clinic. Unfortunately, P. aeruginosa has acquired genes encoding metallo-beta-lactamases (MBLs), enzymes able to hydrolyze most beta-lactam antibiotics. SPM-1 is an MBL produced only by P. aeruginosa, while other MBLs are found in different bacteria. Despite similar active sites, the resistance profile of MBLs towards beta-lactams changes from one enzyme to the other. SPM-1 is unique among pathogen-associated MBLs in that in that it contains "atypical" second sphere residues (S84, G121). Codon randomization on these positions and further selection of resistance-conferring mutants was performed. MICs, periplasmic enzymatic activity, Zn(II) requirements, and protein stability was assessed. Our results indicated that identity of second sphere residues modulates the substrate preferences and the resistance profile of SPM-1 expressed in P. aeruginosa. The second sphere residues found in wild type SPM-1 give rise to a substrate selectivity that is observed only in the periplasmic environment. These residues also allow SPM-1 to confer resistance in P. aeruginosa under Zn(II)-limiting conditions, such as those expected under infection. By optimizing the catalytic efficiency towards beta-lactam antibiotics, the enzyme stability and the Zn(II) binding features, molecular evolution meets the specific needs of a pathogenic bacterial host by means of substitutions outside the active site.Fil: Gonzalez, Lisandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; ArgentinaFil: Bonomo, Robert A.. Case Western Reserve University; Estados UnidosFil: Vila, Alejandro Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaPublic Library Of Science2014-01info: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/11336/7696Gonzalez, Lisandro Javier; Moreno, Diego Martin; Bonomo, Robert A.; Vila, Alejandro Jose; Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues; Public Library Of Science; Plos Pathogens; 10; 1; 1-2014; 1-121553-7366enginfo:eu-repo/semantics/altIdentifier/url/http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3879351/info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.ppat.1003817info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1003817info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:59:16Zoai:ri.conicet.gov.ar:11336/7696instacron: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 09:59:16.722CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
title Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
spellingShingle Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
Gonzalez, Lisandro Javier
Antibiotics
Bacterial Pathogens
Codons
Hydrogen bonding
title_short Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
title_full Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
title_fullStr Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
title_full_unstemmed Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
title_sort Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues
dc.creator.none.fl_str_mv Gonzalez, Lisandro Javier
Moreno, Diego Martin
Bonomo, Robert A.
Vila, Alejandro Jose
author Gonzalez, Lisandro Javier
author_facet Gonzalez, Lisandro Javier
Moreno, Diego Martin
Bonomo, Robert A.
Vila, Alejandro Jose
author_role author
author2 Moreno, Diego Martin
Bonomo, Robert A.
Vila, Alejandro Jose
author2_role author
author
author
dc.subject.none.fl_str_mv Antibiotics
Bacterial Pathogens
Codons
Hydrogen bonding
topic Antibiotics
Bacterial Pathogens
Codons
Hydrogen bonding
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Pseudomonas aeruginosa is one of the most virulent and resistant non-fermenting Gram-negative pathogens in the clinic. Unfortunately, P. aeruginosa has acquired genes encoding metallo-beta-lactamases (MBLs), enzymes able to hydrolyze most beta-lactam antibiotics. SPM-1 is an MBL produced only by P. aeruginosa, while other MBLs are found in different bacteria. Despite similar active sites, the resistance profile of MBLs towards beta-lactams changes from one enzyme to the other. SPM-1 is unique among pathogen-associated MBLs in that in that it contains "atypical" second sphere residues (S84, G121). Codon randomization on these positions and further selection of resistance-conferring mutants was performed. MICs, periplasmic enzymatic activity, Zn(II) requirements, and protein stability was assessed. Our results indicated that identity of second sphere residues modulates the substrate preferences and the resistance profile of SPM-1 expressed in P. aeruginosa. The second sphere residues found in wild type SPM-1 give rise to a substrate selectivity that is observed only in the periplasmic environment. These residues also allow SPM-1 to confer resistance in P. aeruginosa under Zn(II)-limiting conditions, such as those expected under infection. By optimizing the catalytic efficiency towards beta-lactam antibiotics, the enzyme stability and the Zn(II) binding features, molecular evolution meets the specific needs of a pathogenic bacterial host by means of substitutions outside the active site.
Fil: Gonzalez, Lisandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; Argentina
Fil: Bonomo, Robert A.. Case Western Reserve University; Estados Unidos
Fil: Vila, Alejandro Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
description Pseudomonas aeruginosa is one of the most virulent and resistant non-fermenting Gram-negative pathogens in the clinic. Unfortunately, P. aeruginosa has acquired genes encoding metallo-beta-lactamases (MBLs), enzymes able to hydrolyze most beta-lactam antibiotics. SPM-1 is an MBL produced only by P. aeruginosa, while other MBLs are found in different bacteria. Despite similar active sites, the resistance profile of MBLs towards beta-lactams changes from one enzyme to the other. SPM-1 is unique among pathogen-associated MBLs in that in that it contains "atypical" second sphere residues (S84, G121). Codon randomization on these positions and further selection of resistance-conferring mutants was performed. MICs, periplasmic enzymatic activity, Zn(II) requirements, and protein stability was assessed. Our results indicated that identity of second sphere residues modulates the substrate preferences and the resistance profile of SPM-1 expressed in P. aeruginosa. The second sphere residues found in wild type SPM-1 give rise to a substrate selectivity that is observed only in the periplasmic environment. These residues also allow SPM-1 to confer resistance in P. aeruginosa under Zn(II)-limiting conditions, such as those expected under infection. By optimizing the catalytic efficiency towards beta-lactam antibiotics, the enzyme stability and the Zn(II) binding features, molecular evolution meets the specific needs of a pathogenic bacterial host by means of substitutions outside the active site.
publishDate 2014
dc.date.none.fl_str_mv 2014-01
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/11336/7696
Gonzalez, Lisandro Javier; Moreno, Diego Martin; Bonomo, Robert A.; Vila, Alejandro Jose; Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues; Public Library Of Science; Plos Pathogens; 10; 1; 1-2014; 1-12
1553-7366
url http://hdl.handle.net/11336/7696
identifier_str_mv Gonzalez, Lisandro Javier; Moreno, Diego Martin; Bonomo, Robert A.; Vila, Alejandro Jose; Host-Specific Enzyme-Substrate Interactions in SPM-1 Metallo-beta-Lactamase are Modulated by Second Sphere Residues; Public Library Of Science; Plos Pathogens; 10; 1; 1-2014; 1-12
1553-7366
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3879351/
info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.ppat.1003817
info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1003817
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library Of Science
publisher.none.fl_str_mv Public Library Of Science
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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