Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases

Autores
Risso, Valeria Alejandra; Gavira, J. A.; Mejia Carmona, Diego F.; Gaucher, Eric A.; Sanchez Ruiz, José M.
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We report a sequence reconstruction analysis targeting several Precambrian nodes in the evolution of class-A β-lactamases and the preparation and experimental characterization of their encoded proteins. Despite extensive sequence differences with the modern enzymes (100 amino acid differences), the proteins resurrected in the laboratory properly fold into the canonical lactamase structure. The encoded proteins from 2–3 billion years (Gyr)-old β-lactamase sequences undergo cooperative two-state thermal denaturation and display very large denaturation temperature enhancements (35 °C) relative to modern β-lactamases. They degrade different antibiotics in vitro with catalytic efficiencies comparable to that of an average modern enzyme. This enhanced substrate promiscuity is not accompanied by significant changes in the active-site region as seen in static X-ray structures, suggesting a plausible role for dynamics in the evolution of function in these proteins. Laboratory resurrections of 2–3 Gyr-old β-lactamases also endowed modern microorganisms with significant levels of resistance toward a variety of antibiotics, opening up the possibility of performing laboratory replays of the molecular tape of lactamase evolution. Overall, these results support the notions that Precambrian life was thermophilic and that proteins can evolve from substrate-promiscuous generalists into specialists during the course of natural evolution. They also highlight the biotechnological potential of laboratory resurrection of Precambrian proteins, as both high stability and enhanced promiscuity (likely contributors to high evolvability) are advantageous features in protein scaffolds for molecular design and laboratory evolution.
Fil: Risso, Valeria Alejandra. Universidad de Granada. Facultad de Ciencias; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gavira, J. A.. Consejo Superior de Investigaciones Cientificas; España. Universidad de Granada; España
Fil: Mejia Carmona, Diego F.. Universidad de Granada. Facultad de Ciencias; España. Universidad del Valle. Facultad de Salud. Postgrado en Ciencias Biomedicas; Colombia
Fil: Gaucher, Eric A.. Georgia Institute of Techology; Estados Unidos
Fil: Sanchez Ruiz, José M.. Universidad de Granada. Facultad de Ciencias; España
Materia
Ancestral Protein
Hyperstability
Lactamase
Resurrection
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:11336/22624
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamasesRisso, Valeria AlejandraGavira, J. A.Mejia Carmona, Diego F.Gaucher, Eric A.Sanchez Ruiz, José M.Ancestral ProteinHyperstabilityLactamaseResurrectionhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We report a sequence reconstruction analysis targeting several Precambrian nodes in the evolution of class-A β-lactamases and the preparation and experimental characterization of their encoded proteins. Despite extensive sequence differences with the modern enzymes (100 amino acid differences), the proteins resurrected in the laboratory properly fold into the canonical lactamase structure. The encoded proteins from 2–3 billion years (Gyr)-old β-lactamase sequences undergo cooperative two-state thermal denaturation and display very large denaturation temperature enhancements (35 °C) relative to modern β-lactamases. They degrade different antibiotics in vitro with catalytic efficiencies comparable to that of an average modern enzyme. This enhanced substrate promiscuity is not accompanied by significant changes in the active-site region as seen in static X-ray structures, suggesting a plausible role for dynamics in the evolution of function in these proteins. Laboratory resurrections of 2–3 Gyr-old β-lactamases also endowed modern microorganisms with significant levels of resistance toward a variety of antibiotics, opening up the possibility of performing laboratory replays of the molecular tape of lactamase evolution. Overall, these results support the notions that Precambrian life was thermophilic and that proteins can evolve from substrate-promiscuous generalists into specialists during the course of natural evolution. They also highlight the biotechnological potential of laboratory resurrection of Precambrian proteins, as both high stability and enhanced promiscuity (likely contributors to high evolvability) are advantageous features in protein scaffolds for molecular design and laboratory evolution.Fil: Risso, Valeria Alejandra. Universidad de Granada. Facultad de Ciencias; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gavira, J. A.. Consejo Superior de Investigaciones Cientificas; España. Universidad de Granada; EspañaFil: Mejia Carmona, Diego F.. Universidad de Granada. Facultad de Ciencias; España. Universidad del Valle. Facultad de Salud. Postgrado en Ciencias Biomedicas; ColombiaFil: Gaucher, Eric A.. Georgia Institute of Techology; Estados UnidosFil: Sanchez Ruiz, José M.. Universidad de Granada. Facultad de Ciencias; EspañaAmerican Chemical Society2013-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/22624Risso, Valeria Alejandra; Gavira, J. A.; Mejia Carmona, Diego F.; Gaucher, Eric A.; Sanchez Ruiz, José M.; Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases; American Chemical Society; Journal of the American Chemical Society; 135; 8; 2-2013; 2899-29020002-7863CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/abs/10.1021/ja311630ainfo:eu-repo/semantics/altIdentifier/doi/10.1021/ja311630ainfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:58:03Zoai:ri.conicet.gov.ar:11336/22624instacron: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:34982025-10-15 14:58:04.004CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
title Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
spellingShingle Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
Risso, Valeria Alejandra
Ancestral Protein
Hyperstability
Lactamase
Resurrection
title_short Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
title_full Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
title_fullStr Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
title_full_unstemmed Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
title_sort Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases
dc.creator.none.fl_str_mv Risso, Valeria Alejandra
Gavira, J. A.
Mejia Carmona, Diego F.
Gaucher, Eric A.
Sanchez Ruiz, José M.
author Risso, Valeria Alejandra
author_facet Risso, Valeria Alejandra
Gavira, J. A.
Mejia Carmona, Diego F.
Gaucher, Eric A.
Sanchez Ruiz, José M.
author_role author
author2 Gavira, J. A.
Mejia Carmona, Diego F.
Gaucher, Eric A.
Sanchez Ruiz, José M.
author2_role author
author
author
author
dc.subject.none.fl_str_mv Ancestral Protein
Hyperstability
Lactamase
Resurrection
topic Ancestral Protein
Hyperstability
Lactamase
Resurrection
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv We report a sequence reconstruction analysis targeting several Precambrian nodes in the evolution of class-A β-lactamases and the preparation and experimental characterization of their encoded proteins. Despite extensive sequence differences with the modern enzymes (100 amino acid differences), the proteins resurrected in the laboratory properly fold into the canonical lactamase structure. The encoded proteins from 2–3 billion years (Gyr)-old β-lactamase sequences undergo cooperative two-state thermal denaturation and display very large denaturation temperature enhancements (35 °C) relative to modern β-lactamases. They degrade different antibiotics in vitro with catalytic efficiencies comparable to that of an average modern enzyme. This enhanced substrate promiscuity is not accompanied by significant changes in the active-site region as seen in static X-ray structures, suggesting a plausible role for dynamics in the evolution of function in these proteins. Laboratory resurrections of 2–3 Gyr-old β-lactamases also endowed modern microorganisms with significant levels of resistance toward a variety of antibiotics, opening up the possibility of performing laboratory replays of the molecular tape of lactamase evolution. Overall, these results support the notions that Precambrian life was thermophilic and that proteins can evolve from substrate-promiscuous generalists into specialists during the course of natural evolution. They also highlight the biotechnological potential of laboratory resurrection of Precambrian proteins, as both high stability and enhanced promiscuity (likely contributors to high evolvability) are advantageous features in protein scaffolds for molecular design and laboratory evolution.
Fil: Risso, Valeria Alejandra. Universidad de Granada. Facultad de Ciencias; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gavira, J. A.. Consejo Superior de Investigaciones Cientificas; España. Universidad de Granada; España
Fil: Mejia Carmona, Diego F.. Universidad de Granada. Facultad de Ciencias; España. Universidad del Valle. Facultad de Salud. Postgrado en Ciencias Biomedicas; Colombia
Fil: Gaucher, Eric A.. Georgia Institute of Techology; Estados Unidos
Fil: Sanchez Ruiz, José M.. Universidad de Granada. Facultad de Ciencias; España
description We report a sequence reconstruction analysis targeting several Precambrian nodes in the evolution of class-A β-lactamases and the preparation and experimental characterization of their encoded proteins. Despite extensive sequence differences with the modern enzymes (100 amino acid differences), the proteins resurrected in the laboratory properly fold into the canonical lactamase structure. The encoded proteins from 2–3 billion years (Gyr)-old β-lactamase sequences undergo cooperative two-state thermal denaturation and display very large denaturation temperature enhancements (35 °C) relative to modern β-lactamases. They degrade different antibiotics in vitro with catalytic efficiencies comparable to that of an average modern enzyme. This enhanced substrate promiscuity is not accompanied by significant changes in the active-site region as seen in static X-ray structures, suggesting a plausible role for dynamics in the evolution of function in these proteins. Laboratory resurrections of 2–3 Gyr-old β-lactamases also endowed modern microorganisms with significant levels of resistance toward a variety of antibiotics, opening up the possibility of performing laboratory replays of the molecular tape of lactamase evolution. Overall, these results support the notions that Precambrian life was thermophilic and that proteins can evolve from substrate-promiscuous generalists into specialists during the course of natural evolution. They also highlight the biotechnological potential of laboratory resurrection of Precambrian proteins, as both high stability and enhanced promiscuity (likely contributors to high evolvability) are advantageous features in protein scaffolds for molecular design and laboratory evolution.
publishDate 2013
dc.date.none.fl_str_mv 2013-02
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/11336/22624
Risso, Valeria Alejandra; Gavira, J. A.; Mejia Carmona, Diego F.; Gaucher, Eric A.; Sanchez Ruiz, José M.; Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases; American Chemical Society; Journal of the American Chemical Society; 135; 8; 2-2013; 2899-2902
0002-7863
CONICET Digital
CONICET
url http://hdl.handle.net/11336/22624
identifier_str_mv Risso, Valeria Alejandra; Gavira, J. A.; Mejia Carmona, Diego F.; Gaucher, Eric A.; Sanchez Ruiz, José M.; Hyperstability and substrate promiscuity in laboratory resurrections of precambrian β-lactamases; American Chemical Society; Journal of the American Chemical Society; 135; 8; 2-2013; 2899-2902
0002-7863
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/abs/10.1021/ja311630a
info:eu-repo/semantics/altIdentifier/doi/10.1021/ja311630a
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
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
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 13.22299

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