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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/7696
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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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1844613760046596096 |
score |
13.070432 |