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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/93960
Ver los metadatos del registro completo
id |
CONICETDig_7ee34752ed75d85d08df3c7d65c5fc36 |
---|---|
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 Técnicas2025-09-29T10:21:33Zoai: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:34982025-09-29 10:21:33.474CONICET 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 |
_version_ |
1844614204460367872 |
score |
13.070432 |