Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase

Autores
Gonzalez, Lisandro Javier; Stival, Cintia Estefanía; Puzzolo, Juan Luis; Moreno, Diego Martin; Aguilar Vila, Alejandro
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Metallo-β-lactamases (MBLs) are the major group of carbapenemases produced by bacterial pathogens. The design of MBL inhibitors has been limited by, among other issues, incomplete knowledge about how these enzymes modulate substrate recognition. While most MBLs are broad-spectrum enzymes, B2 MBLs are exclusive carbapenemases. This narrower substrate profile has been attributed to a sequence insertion present in B2 enzymes that limits accessibility to the active site. In this work, we evaluate the role of sequence insertions naturally occurring in the B2 enzyme Sfh-I and in the broad-spectrum B1 enzyme SPM-1. We engineered a chimeric protein in which the sequence insertion of SPM-1 was replaced by the one present in Sfh-I. The chimeric variant is a selective cephalosporinase, revealing that the substrate profile of MBLs can be further tuned depending on the protein context. These results also show that the stable scaffold of MBLs allows a modular engineering much richer than the one observed in nature.
Fil: González, Lisandro J.. Instituto de Biologia Molecular y Celular de Rosario; Argentina
Fil: Stival, Cintia Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Puzzolo, Juan Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina
Fil: Moreno, Diego M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina
Fil: Aguilar Vila, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Materia
BETA-LACTAMASES
MECHANISMS OF RESISTANCE
METALLO-BETA-LACTAMASE
SPM-1
SUBSTRATE PROFILE
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:11336/93960
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/93960
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Shaping substrate selectivity in a broad-spectrum metallo-β-lactamaseGonzalez, Lisandro JavierStival, Cintia EstefaníaPuzzolo, Juan LuisMoreno, Diego MartinAguilar Vila, AlejandroBETA-LACTAMASESMECHANISMS OF RESISTANCEMETALLO-BETA-LACTAMASESPM-1SUBSTRATE PROFILEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Metallo-β-lactamases (MBLs) are the major group of carbapenemases produced by bacterial pathogens. The design of MBL inhibitors has been limited by, among other issues, incomplete knowledge about how these enzymes modulate substrate recognition. While most MBLs are broad-spectrum enzymes, B2 MBLs are exclusive carbapenemases. This narrower substrate profile has been attributed to a sequence insertion present in B2 enzymes that limits accessibility to the active site. In this work, we evaluate the role of sequence insertions naturally occurring in the B2 enzyme Sfh-I and in the broad-spectrum B1 enzyme SPM-1. We engineered a chimeric protein in which the sequence insertion of SPM-1 was replaced by the one present in Sfh-I. The chimeric variant is a selective cephalosporinase, revealing that the substrate profile of MBLs can be further tuned depending on the protein context. These results also show that the stable scaffold of MBLs allows a modular engineering much richer than the one observed in nature.Fil: González, Lisandro J.. Instituto de Biologia Molecular y Celular de Rosario; ArgentinaFil: Stival, Cintia Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Puzzolo, Juan Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; ArgentinaFil: Moreno, Diego M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; ArgentinaFil: Aguilar Vila, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaAmerican Society for Microbiology2018-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/93960Gonzalez, Lisandro Javier; Stival, Cintia Estefanía; Puzzolo, Juan Luis; Moreno, Diego Martin; Aguilar Vila, Alejandro; Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase; American Society for Microbiology; Antimicrobial Agents and Chemotherapy; 62; 4; 4-2018; 1-300066-4804CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://aac.asm.org/content/62/4/e02079-17info:eu-repo/semantics/altIdentifier/doi/10.1128/AAC.02079-17info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:54:19Zoai:ri.conicet.gov.ar:11336/93960instacron: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:34982025-10-22 11:54:19.759CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
title Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
spellingShingle Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
Gonzalez, Lisandro Javier
BETA-LACTAMASES
MECHANISMS OF RESISTANCE
METALLO-BETA-LACTAMASE
SPM-1
SUBSTRATE PROFILE
title_short Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
title_full Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
title_fullStr Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
title_full_unstemmed Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
title_sort Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase
dc.creator.none.fl_str_mv Gonzalez, Lisandro Javier
Stival, Cintia Estefanía
Puzzolo, Juan Luis
Moreno, Diego Martin
Aguilar Vila, Alejandro
author Gonzalez, Lisandro Javier
author_facet Gonzalez, Lisandro Javier
Stival, Cintia Estefanía
Puzzolo, Juan Luis
Moreno, Diego Martin
Aguilar Vila, Alejandro
author_role author
author2 Stival, Cintia Estefanía
Puzzolo, Juan Luis
Moreno, Diego Martin
Aguilar Vila, Alejandro
author2_role author
author
author
author
dc.subject.none.fl_str_mv BETA-LACTAMASES
MECHANISMS OF RESISTANCE
METALLO-BETA-LACTAMASE
SPM-1
SUBSTRATE PROFILE
topic BETA-LACTAMASES
MECHANISMS OF RESISTANCE
METALLO-BETA-LACTAMASE
SPM-1
SUBSTRATE PROFILE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Metallo-β-lactamases (MBLs) are the major group of carbapenemases produced by bacterial pathogens. The design of MBL inhibitors has been limited by, among other issues, incomplete knowledge about how these enzymes modulate substrate recognition. While most MBLs are broad-spectrum enzymes, B2 MBLs are exclusive carbapenemases. This narrower substrate profile has been attributed to a sequence insertion present in B2 enzymes that limits accessibility to the active site. In this work, we evaluate the role of sequence insertions naturally occurring in the B2 enzyme Sfh-I and in the broad-spectrum B1 enzyme SPM-1. We engineered a chimeric protein in which the sequence insertion of SPM-1 was replaced by the one present in Sfh-I. The chimeric variant is a selective cephalosporinase, revealing that the substrate profile of MBLs can be further tuned depending on the protein context. These results also show that the stable scaffold of MBLs allows a modular engineering much richer than the one observed in nature.
Fil: González, Lisandro J.. Instituto de Biologia Molecular y Celular de Rosario; Argentina
Fil: Stival, Cintia Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Puzzolo, Juan Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina
Fil: Moreno, Diego M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina
Fil: Aguilar Vila, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
description Metallo-β-lactamases (MBLs) are the major group of carbapenemases produced by bacterial pathogens. The design of MBL inhibitors has been limited by, among other issues, incomplete knowledge about how these enzymes modulate substrate recognition. While most MBLs are broad-spectrum enzymes, B2 MBLs are exclusive carbapenemases. This narrower substrate profile has been attributed to a sequence insertion present in B2 enzymes that limits accessibility to the active site. In this work, we evaluate the role of sequence insertions naturally occurring in the B2 enzyme Sfh-I and in the broad-spectrum B1 enzyme SPM-1. We engineered a chimeric protein in which the sequence insertion of SPM-1 was replaced by the one present in Sfh-I. The chimeric variant is a selective cephalosporinase, revealing that the substrate profile of MBLs can be further tuned depending on the protein context. These results also show that the stable scaffold of MBLs allows a modular engineering much richer than the one observed in nature.
publishDate 2018
dc.date.none.fl_str_mv 2018-04
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/11336/93960
Gonzalez, Lisandro Javier; Stival, Cintia Estefanía; Puzzolo, Juan Luis; Moreno, Diego Martin; Aguilar Vila, Alejandro; Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase; American Society for Microbiology; Antimicrobial Agents and Chemotherapy; 62; 4; 4-2018; 1-30
0066-4804
CONICET Digital
CONICET
url http://hdl.handle.net/11336/93960
identifier_str_mv Gonzalez, Lisandro Javier; Stival, Cintia Estefanía; Puzzolo, Juan Luis; Moreno, Diego Martin; Aguilar Vila, Alejandro; Shaping substrate selectivity in a broad-spectrum metallo-β-lactamase; American Society for Microbiology; Antimicrobial Agents and Chemotherapy; 62; 4; 4-2018; 1-30
0066-4804
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://aac.asm.org/content/62/4/e02079-17
info:eu-repo/semantics/altIdentifier/doi/10.1128/AAC.02079-17
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
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 12.982451

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