Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase

Autores
Ruggiero, Melina; Papp Wallace, Krisztina M.; Taracila, Magdalena A.; Mojica, Maria F.; Bethel, Christopher R.; Rudin, Susan D.; Zeiser, Elise T.; Gutkind, Gabriel Osvaldo; Bonomo, Robert A.; Power, Pablo
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
PER β -lactamases are an emerging family of extended-spectrum β -lactamases (ESBL) found in Gram-negative bacteria. PER β -lactamases are unique among class A enzymes as they possess an inverted omega (?) loop and extended B3 β -strand. These singular structural features are hypothesized to contribute to their hydrolytic profile against oxyimino-cephalosporins (e.g., cefotaxime and ceftazidime). Here, we tested the ability of avibactam (AVI), a novel non- β -lactam β -lactamase inhibitor to inactivate PER-2. Interestingly, the PER-2 inhibition constants (i.e., k2/K = 2 × 103 ±0.1 × 103 M-1 s-1, where k2 is the rate constant for acylation (carbamylation) and K is the equilibrium constant) that were obtained when AVI was tested were reminiscent of values observed testing the inhibition by AVI of class C and D β -lactamases (i.e., k2/K range of =103 M-1s-1) and not class A β -lactamases (i.e., k2/K range, 104 to 105 M-1s-1). Once AVI was bound, a stable complex with PER-2 was observed via mass spectrometry (e.g., 31,389 ± 3 atomic mass units [amu] ¡ 31,604 ± 3 amu for 24 h). Molecular modeling of PER-2 with AVI showed that the carbonyl of AVI was located in the oxyanion hole of the β -lactamase and that the sulfate of AVI formed interactions with the β-lactam carboxylate binding site of the PER-2 β -lactamase (R220 and T237). However, hydrophobic patches near the PER-2 active site (by Ser70 and B3-B4 β -strands) were observed and may affect the binding of necessary catalytic water molecules, thus slowing acylation (k2/K) of AVI onto PER-2. Similar electrostatics and hydrophobicity of the active site were also observed between OXA-48 and PER-2, while CTX-M-15 was more hydrophilic. To demonstrate the ability of AVI to overcome the enhanced cephalosporinase activity of PER-2 β-lactamase, we tested different β -lactam-AVI combinations. By lowering MICs to ≤2 mg/liter, the ceftaroline-AVI combination could represent a favorable therapeutic option against Enterobacteriaceae expressing blaPER-2. Our studies define the inactivation of the PER-2 ESBL by AVI and suggest that the biophysical properties of the active site contribute to determining the efficiency of inactivation.
Fil: Ruggiero, Melina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina
Fil: Papp Wallace, Krisztina M.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Taracila, Magdalena A.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Mojica, Maria F.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos
Fil: Bethel, Christopher R.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos
Fil: Rudin, Susan D.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Zeiser, Elise T.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos
Fil: Gutkind, Gabriel Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina
Fil: Bonomo, Robert A.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Power, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina
Materia
AVIBACTAM
BETA-LACTAMASES
BETA-LACTAMS
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:11336/48832
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/48832
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-LactamaseRuggiero, MelinaPapp Wallace, Krisztina M.Taracila, Magdalena A.Mojica, Maria F.Bethel, Christopher R.Rudin, Susan D.Zeiser, Elise T.Gutkind, Gabriel OsvaldoBonomo, Robert A.Power, PabloAVIBACTAMBETA-LACTAMASESBETA-LACTAMShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1PER β -lactamases are an emerging family of extended-spectrum β -lactamases (ESBL) found in Gram-negative bacteria. PER β -lactamases are unique among class A enzymes as they possess an inverted omega (?) loop and extended B3 β -strand. These singular structural features are hypothesized to contribute to their hydrolytic profile against oxyimino-cephalosporins (e.g., cefotaxime and ceftazidime). Here, we tested the ability of avibactam (AVI), a novel non- β -lactam β -lactamase inhibitor to inactivate PER-2. Interestingly, the PER-2 inhibition constants (i.e., k2/K = 2 × 103 ±0.1 × 103 M-1 s-1, where k2 is the rate constant for acylation (carbamylation) and K is the equilibrium constant) that were obtained when AVI was tested were reminiscent of values observed testing the inhibition by AVI of class C and D β -lactamases (i.e., k2/K range of =103 M-1s-1) and not class A β -lactamases (i.e., k2/K range, 104 to 105 M-1s-1). Once AVI was bound, a stable complex with PER-2 was observed via mass spectrometry (e.g., 31,389 ± 3 atomic mass units [amu] ¡ 31,604 ± 3 amu for 24 h). Molecular modeling of PER-2 with AVI showed that the carbonyl of AVI was located in the oxyanion hole of the β -lactamase and that the sulfate of AVI formed interactions with the β-lactam carboxylate binding site of the PER-2 β -lactamase (R220 and T237). However, hydrophobic patches near the PER-2 active site (by Ser70 and B3-B4 β -strands) were observed and may affect the binding of necessary catalytic water molecules, thus slowing acylation (k2/K) of AVI onto PER-2. Similar electrostatics and hydrophobicity of the active site were also observed between OXA-48 and PER-2, while CTX-M-15 was more hydrophilic. To demonstrate the ability of AVI to overcome the enhanced cephalosporinase activity of PER-2 β-lactamase, we tested different β -lactam-AVI combinations. By lowering MICs to ≤2 mg/liter, the ceftaroline-AVI combination could represent a favorable therapeutic option against Enterobacteriaceae expressing blaPER-2. Our studies define the inactivation of the PER-2 ESBL by AVI and suggest that the biophysical properties of the active site contribute to determining the efficiency of inactivation.Fil: Ruggiero, Melina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; ArgentinaFil: Papp Wallace, Krisztina M.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados UnidosFil: Taracila, Magdalena A.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados UnidosFil: Mojica, Maria F.. Louis Stokes Cleveland Department of Veterans Affairs; Estados UnidosFil: Bethel, Christopher R.. Louis Stokes Cleveland Department of Veterans Affairs; Estados UnidosFil: Rudin, Susan D.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados UnidosFil: Zeiser, Elise T.. Louis Stokes Cleveland Department of Veterans Affairs; Estados UnidosFil: Gutkind, Gabriel Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; ArgentinaFil: Bonomo, Robert A.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados UnidosFil: Power, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; ArgentinaAmerican Society for Microbiology2017-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/48832Ruggiero, Melina; Papp Wallace, Krisztina M.; Taracila, Magdalena A.; Mojica, Maria F.; Bethel, Christopher R.; et al.; Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase; American Society for Microbiology; Antimicrobial Agents and Chemotherapy; 61; 6; 6-20170066-4804CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1128/AAC.02476-16info:eu-repo/semantics/altIdentifier/url/http://aac.asm.org/content/61/6/e02476-16info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:44:53Zoai:ri.conicet.gov.ar:11336/48832instacron: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 15:44:53.543CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
title Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
spellingShingle Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
Ruggiero, Melina
AVIBACTAM
BETA-LACTAMASES
BETA-LACTAMS
title_short Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
title_full Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
title_fullStr Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
title_full_unstemmed Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
title_sort Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase
dc.creator.none.fl_str_mv Ruggiero, Melina
Papp Wallace, Krisztina M.
Taracila, Magdalena A.
Mojica, Maria F.
Bethel, Christopher R.
Rudin, Susan D.
Zeiser, Elise T.
Gutkind, Gabriel Osvaldo
Bonomo, Robert A.
Power, Pablo
author Ruggiero, Melina
author_facet Ruggiero, Melina
Papp Wallace, Krisztina M.
Taracila, Magdalena A.
Mojica, Maria F.
Bethel, Christopher R.
Rudin, Susan D.
Zeiser, Elise T.
Gutkind, Gabriel Osvaldo
Bonomo, Robert A.
Power, Pablo
author_role author
author2 Papp Wallace, Krisztina M.
Taracila, Magdalena A.
Mojica, Maria F.
Bethel, Christopher R.
Rudin, Susan D.
Zeiser, Elise T.
Gutkind, Gabriel Osvaldo
Bonomo, Robert A.
Power, Pablo
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv AVIBACTAM
BETA-LACTAMASES
BETA-LACTAMS
topic AVIBACTAM
BETA-LACTAMASES
BETA-LACTAMS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv PER β -lactamases are an emerging family of extended-spectrum β -lactamases (ESBL) found in Gram-negative bacteria. PER β -lactamases are unique among class A enzymes as they possess an inverted omega (?) loop and extended B3 β -strand. These singular structural features are hypothesized to contribute to their hydrolytic profile against oxyimino-cephalosporins (e.g., cefotaxime and ceftazidime). Here, we tested the ability of avibactam (AVI), a novel non- β -lactam β -lactamase inhibitor to inactivate PER-2. Interestingly, the PER-2 inhibition constants (i.e., k2/K = 2 × 103 ±0.1 × 103 M-1 s-1, where k2 is the rate constant for acylation (carbamylation) and K is the equilibrium constant) that were obtained when AVI was tested were reminiscent of values observed testing the inhibition by AVI of class C and D β -lactamases (i.e., k2/K range of =103 M-1s-1) and not class A β -lactamases (i.e., k2/K range, 104 to 105 M-1s-1). Once AVI was bound, a stable complex with PER-2 was observed via mass spectrometry (e.g., 31,389 ± 3 atomic mass units [amu] ¡ 31,604 ± 3 amu for 24 h). Molecular modeling of PER-2 with AVI showed that the carbonyl of AVI was located in the oxyanion hole of the β -lactamase and that the sulfate of AVI formed interactions with the β-lactam carboxylate binding site of the PER-2 β -lactamase (R220 and T237). However, hydrophobic patches near the PER-2 active site (by Ser70 and B3-B4 β -strands) were observed and may affect the binding of necessary catalytic water molecules, thus slowing acylation (k2/K) of AVI onto PER-2. Similar electrostatics and hydrophobicity of the active site were also observed between OXA-48 and PER-2, while CTX-M-15 was more hydrophilic. To demonstrate the ability of AVI to overcome the enhanced cephalosporinase activity of PER-2 β-lactamase, we tested different β -lactam-AVI combinations. By lowering MICs to ≤2 mg/liter, the ceftaroline-AVI combination could represent a favorable therapeutic option against Enterobacteriaceae expressing blaPER-2. Our studies define the inactivation of the PER-2 ESBL by AVI and suggest that the biophysical properties of the active site contribute to determining the efficiency of inactivation.
Fil: Ruggiero, Melina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina
Fil: Papp Wallace, Krisztina M.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Taracila, Magdalena A.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Mojica, Maria F.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos
Fil: Bethel, Christopher R.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos
Fil: Rudin, Susan D.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Zeiser, Elise T.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos
Fil: Gutkind, Gabriel Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina
Fil: Bonomo, Robert A.. Louis Stokes Cleveland Department of Veterans Affairs; Estados Unidos. Case Western Reserve University; Estados Unidos
Fil: Power, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina
description PER β -lactamases are an emerging family of extended-spectrum β -lactamases (ESBL) found in Gram-negative bacteria. PER β -lactamases are unique among class A enzymes as they possess an inverted omega (?) loop and extended B3 β -strand. These singular structural features are hypothesized to contribute to their hydrolytic profile against oxyimino-cephalosporins (e.g., cefotaxime and ceftazidime). Here, we tested the ability of avibactam (AVI), a novel non- β -lactam β -lactamase inhibitor to inactivate PER-2. Interestingly, the PER-2 inhibition constants (i.e., k2/K = 2 × 103 ±0.1 × 103 M-1 s-1, where k2 is the rate constant for acylation (carbamylation) and K is the equilibrium constant) that were obtained when AVI was tested were reminiscent of values observed testing the inhibition by AVI of class C and D β -lactamases (i.e., k2/K range of =103 M-1s-1) and not class A β -lactamases (i.e., k2/K range, 104 to 105 M-1s-1). Once AVI was bound, a stable complex with PER-2 was observed via mass spectrometry (e.g., 31,389 ± 3 atomic mass units [amu] ¡ 31,604 ± 3 amu for 24 h). Molecular modeling of PER-2 with AVI showed that the carbonyl of AVI was located in the oxyanion hole of the β -lactamase and that the sulfate of AVI formed interactions with the β-lactam carboxylate binding site of the PER-2 β -lactamase (R220 and T237). However, hydrophobic patches near the PER-2 active site (by Ser70 and B3-B4 β -strands) were observed and may affect the binding of necessary catalytic water molecules, thus slowing acylation (k2/K) of AVI onto PER-2. Similar electrostatics and hydrophobicity of the active site were also observed between OXA-48 and PER-2, while CTX-M-15 was more hydrophilic. To demonstrate the ability of AVI to overcome the enhanced cephalosporinase activity of PER-2 β-lactamase, we tested different β -lactam-AVI combinations. By lowering MICs to ≤2 mg/liter, the ceftaroline-AVI combination could represent a favorable therapeutic option against Enterobacteriaceae expressing blaPER-2. Our studies define the inactivation of the PER-2 ESBL by AVI and suggest that the biophysical properties of the active site contribute to determining the efficiency of inactivation.
publishDate 2017
dc.date.none.fl_str_mv 2017-06
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/11336/48832
Ruggiero, Melina; Papp Wallace, Krisztina M.; Taracila, Magdalena A.; Mojica, Maria F.; Bethel, Christopher R.; et al.; Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase; American Society for Microbiology; Antimicrobial Agents and Chemotherapy; 61; 6; 6-2017
0066-4804
CONICET Digital
CONICET
url http://hdl.handle.net/11336/48832
identifier_str_mv Ruggiero, Melina; Papp Wallace, Krisztina M.; Taracila, Magdalena A.; Mojica, Maria F.; Bethel, Christopher R.; et al.; Exploring the landscape of diazabicyclooctane (DBO) inhibition: Avibactam inactivation of PER-2 β-Lactamase; American Society for Microbiology; Antimicrobial Agents and Chemotherapy; 61; 6; 6-2017
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/doi/10.1128/AAC.02476-16
info:eu-repo/semantics/altIdentifier/url/http://aac.asm.org/content/61/6/e02476-16
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
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 13.221938

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