Optical studies of single-tryptophan B. licheniformis β-lactamase variants

Autores
Risso, Valeria Alejandra; Primo, Maria Evangelina; Brunet, Juan E.; Sotomayor, Carlos P.; Ermacora, Mario Roberto
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
β-lactamases (penicillinases) are important complicating factors in bacterial infections and excellent theoretical and experimental models in protein structure, dynamics and evolution. Bacillus licheniformis exo-small penicillinase (ESP) is a Class A β-lactamase with three tryptophan residues, one located in each of the two protein domains and one located in the interface between domains. To determine the tryptophan contribution to the ESP UV-absorption, circular dichroism, and steady-state and time-resolved fluorescence, four Trp. →. Phe mutants were prepared and characterized. The residue substitutions had little impact on the native conformation. UV-absorption and CD features were identified and ascribed to specific aromatic residues. Time-resolved fluorescence showed that most of the fluorescence decay of ESP tryptophans is due to a discrete exponential component with a lifetime of 5-6. ns. Fluorescence polarization measurements indicated that fluorescence of Trp 210 is nearly independent of the fluorescence of Trp 229 and Trp 251, whereas a substantial energy homotransfer between the latter pair takes place. The spectroscopic information was rationalized on the basis of structural considerations and should help in the interpretation and monitoring of the changes at the sub domain level during the conformational transitions and fluctuations of ESP and other Class A β-lactamases.
Fil: Risso, Valeria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina
Fil: Primo, Maria Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; Argentina
Fil: Brunet, Juan E.. Pontificia Universidad Católica de Valparaíso; Chile
Fil: Sotomayor, Carlos P.. Pontificia Universidad Católica de Valparaíso; Chile
Fil: Ermacora, Mario Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina
Materia
Β-LACTAMASE
CIRCULAR DICHROISM
PROTEIN CONFORMATION
TRYPTOPHAN FLUORESCENCE
UV-ABSORPTION
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:11336/97942
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/97942
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Optical studies of single-tryptophan B. licheniformis β-lactamase variantsRisso, Valeria AlejandraPrimo, Maria EvangelinaBrunet, Juan E.Sotomayor, Carlos P.Ermacora, Mario RobertoΒ-LACTAMASECIRCULAR DICHROISMPROTEIN CONFORMATIONTRYPTOPHAN FLUORESCENCEUV-ABSORPTIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1β-lactamases (penicillinases) are important complicating factors in bacterial infections and excellent theoretical and experimental models in protein structure, dynamics and evolution. Bacillus licheniformis exo-small penicillinase (ESP) is a Class A β-lactamase with three tryptophan residues, one located in each of the two protein domains and one located in the interface between domains. To determine the tryptophan contribution to the ESP UV-absorption, circular dichroism, and steady-state and time-resolved fluorescence, four Trp. →. Phe mutants were prepared and characterized. The residue substitutions had little impact on the native conformation. UV-absorption and CD features were identified and ascribed to specific aromatic residues. Time-resolved fluorescence showed that most of the fluorescence decay of ESP tryptophans is due to a discrete exponential component with a lifetime of 5-6. ns. Fluorescence polarization measurements indicated that fluorescence of Trp 210 is nearly independent of the fluorescence of Trp 229 and Trp 251, whereas a substantial energy homotransfer between the latter pair takes place. The spectroscopic information was rationalized on the basis of structural considerations and should help in the interpretation and monitoring of the changes at the sub domain level during the conformational transitions and fluctuations of ESP and other Class A β-lactamases.Fil: Risso, Valeria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; ArgentinaFil: Primo, Maria Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Brunet, Juan E.. Pontificia Universidad Católica de Valparaíso; ChileFil: Sotomayor, Carlos P.. Pontificia Universidad Católica de Valparaíso; ChileFil: Ermacora, Mario Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; ArgentinaElsevier Science2010-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/97942Risso, Valeria Alejandra; Primo, Maria Evangelina; Brunet, Juan E.; Sotomayor, Carlos P.; Ermacora, Mario Roberto; Optical studies of single-tryptophan B. licheniformis β-lactamase variants; Elsevier Science; Biophysical Chemistry; 151; 3; 10-2010; 111-1180301-4622CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0301462210001560info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpc.2010.05.013info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:00:41Zoai:ri.conicet.gov.ar:11336/97942instacron: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 10:00:41.45CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optical studies of single-tryptophan B. licheniformis β-lactamase variants
title Optical studies of single-tryptophan B. licheniformis β-lactamase variants
spellingShingle Optical studies of single-tryptophan B. licheniformis β-lactamase variants
Risso, Valeria Alejandra
Β-LACTAMASE
CIRCULAR DICHROISM
PROTEIN CONFORMATION
TRYPTOPHAN FLUORESCENCE
UV-ABSORPTION
title_short Optical studies of single-tryptophan B. licheniformis β-lactamase variants
title_full Optical studies of single-tryptophan B. licheniformis β-lactamase variants
title_fullStr Optical studies of single-tryptophan B. licheniformis β-lactamase variants
title_full_unstemmed Optical studies of single-tryptophan B. licheniformis β-lactamase variants
title_sort Optical studies of single-tryptophan B. licheniformis β-lactamase variants
dc.creator.none.fl_str_mv Risso, Valeria Alejandra
Primo, Maria Evangelina
Brunet, Juan E.
Sotomayor, Carlos P.
Ermacora, Mario Roberto
author Risso, Valeria Alejandra
author_facet Risso, Valeria Alejandra
Primo, Maria Evangelina
Brunet, Juan E.
Sotomayor, Carlos P.
Ermacora, Mario Roberto
author_role author
author2 Primo, Maria Evangelina
Brunet, Juan E.
Sotomayor, Carlos P.
Ermacora, Mario Roberto
author2_role author
author
author
author
dc.subject.none.fl_str_mv Β-LACTAMASE
CIRCULAR DICHROISM
PROTEIN CONFORMATION
TRYPTOPHAN FLUORESCENCE
UV-ABSORPTION
topic Β-LACTAMASE
CIRCULAR DICHROISM
PROTEIN CONFORMATION
TRYPTOPHAN FLUORESCENCE
UV-ABSORPTION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv β-lactamases (penicillinases) are important complicating factors in bacterial infections and excellent theoretical and experimental models in protein structure, dynamics and evolution. Bacillus licheniformis exo-small penicillinase (ESP) is a Class A β-lactamase with three tryptophan residues, one located in each of the two protein domains and one located in the interface between domains. To determine the tryptophan contribution to the ESP UV-absorption, circular dichroism, and steady-state and time-resolved fluorescence, four Trp. →. Phe mutants were prepared and characterized. The residue substitutions had little impact on the native conformation. UV-absorption and CD features were identified and ascribed to specific aromatic residues. Time-resolved fluorescence showed that most of the fluorescence decay of ESP tryptophans is due to a discrete exponential component with a lifetime of 5-6. ns. Fluorescence polarization measurements indicated that fluorescence of Trp 210 is nearly independent of the fluorescence of Trp 229 and Trp 251, whereas a substantial energy homotransfer between the latter pair takes place. The spectroscopic information was rationalized on the basis of structural considerations and should help in the interpretation and monitoring of the changes at the sub domain level during the conformational transitions and fluctuations of ESP and other Class A β-lactamases.
Fil: Risso, Valeria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina
Fil: Primo, Maria Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; Argentina
Fil: Brunet, Juan E.. Pontificia Universidad Católica de Valparaíso; Chile
Fil: Sotomayor, Carlos P.. Pontificia Universidad Católica de Valparaíso; Chile
Fil: Ermacora, Mario Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina
description β-lactamases (penicillinases) are important complicating factors in bacterial infections and excellent theoretical and experimental models in protein structure, dynamics and evolution. Bacillus licheniformis exo-small penicillinase (ESP) is a Class A β-lactamase with three tryptophan residues, one located in each of the two protein domains and one located in the interface between domains. To determine the tryptophan contribution to the ESP UV-absorption, circular dichroism, and steady-state and time-resolved fluorescence, four Trp. →. Phe mutants were prepared and characterized. The residue substitutions had little impact on the native conformation. UV-absorption and CD features were identified and ascribed to specific aromatic residues. Time-resolved fluorescence showed that most of the fluorescence decay of ESP tryptophans is due to a discrete exponential component with a lifetime of 5-6. ns. Fluorescence polarization measurements indicated that fluorescence of Trp 210 is nearly independent of the fluorescence of Trp 229 and Trp 251, whereas a substantial energy homotransfer between the latter pair takes place. The spectroscopic information was rationalized on the basis of structural considerations and should help in the interpretation and monitoring of the changes at the sub domain level during the conformational transitions and fluctuations of ESP and other Class A β-lactamases.
publishDate 2010
dc.date.none.fl_str_mv 2010-10
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/11336/97942
Risso, Valeria Alejandra; Primo, Maria Evangelina; Brunet, Juan E.; Sotomayor, Carlos P.; Ermacora, Mario Roberto; Optical studies of single-tryptophan B. licheniformis β-lactamase variants; Elsevier Science; Biophysical Chemistry; 151; 3; 10-2010; 111-118
0301-4622
CONICET Digital
CONICET
url http://hdl.handle.net/11336/97942
identifier_str_mv Risso, Valeria Alejandra; Primo, Maria Evangelina; Brunet, Juan E.; Sotomayor, Carlos P.; Ermacora, Mario Roberto; Optical studies of single-tryptophan B. licheniformis β-lactamase variants; Elsevier Science; Biophysical Chemistry; 151; 3; 10-2010; 111-118
0301-4622
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://www.sciencedirect.com/science/article/pii/S0301462210001560
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpc.2010.05.013
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
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier 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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score 13.070432

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