Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase

Autores
Souza, Tatiana A. C. B.; Okamoto, Débora N.; Ruiz, Diego M.; Oliveira, Lilian C. G.; Kondo, Márcia Y.; Tersario, Ivarne L. S.; Juliano, Luiz; de Castro, Rosana Esther; Gouvea, Iuri E.; Murakami, Mário T.
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Nep (Natrialba magadii extracellular protease) is a halolysin-like peptidase secreted by the haloalkaliphilic archaeon N. magadii that exhibits optimal activity and stability in salt-saturated solutions. In this work, the effect of salt on the function and structure of Nep was investigated. In absence of salt, Nep became unfolded and aggregated, leading to the loss of activity. The enzyme did not recover its structural and functional properties even after restoring the ideal conditions for catalysis. At salt concentrations higher than 1 M (NaCl), Nep behaved as monomers in solution and its enzymatic activity displayed a nonlinear concave-up dependence with salt concentration resulting in a 20-fold activation at 4 M NaCl. Although transition from a high to a low-saline environment (3–1 M NaCl) did not affect its secondary structure contents, it diminished the enzyme stability and provoked large structural rearrangements, changing from an elongated shape at 3 M NaCl to a compact conformational state at 1 M NaCl. The thermodynamic analysis of peptide hydrolysis by Nep suggests a significant enzyme reorganization depending on the environmental salinity, which supports in solution SAXS and DLS studies. Moreover, solvent kinetic isotopic effect (SKIE) data indicates the general acid-base mechanism as the rate-limiting step for Nep catalysis, like classical serine-peptidases. All these data correlate the Nep conformational states with the enzymatic behavior providing a further understanding on the stability and structural determinants for the functioning of halolysins under different salinities.
Fil: Souza, Tatiana A. C. B.. Centro Nacional de Pesquisas em Energia e Materiais; Brasil
Fil: Okamoto, Débora N.. Universidade de Sao Paulo; Brasil
Fil: Ruiz, Diego M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Oliveira, Lilian C. G.. Universidade de Sao Paulo; Brasil
Fil: Kondo, Márcia Y.. Universidade de Sao Paulo; Brasil
Fil: Tersario, Ivarne L. S.. Universidade de Mogi das Cruzes; Brasil
Fil: Juliano, Luiz. Universidade de Sao Paulo; Brasil
Fil: de Castro, Rosana Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Gouvea, Iuri E.. Universidade de Sao Paulo; Brasil
Fil: Murakami, Mário T.. Centro Nacional de Pesquisas em Energia e Materiais; Brasil
Materia
Natrialba Magadii
Extracellular Protease
Halophilism
Structure
Stability
Kinetics
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:11336/13119
Acceder
id CONICETDig_b7b0e8e3bd000a28e26892f5524e5b39
oai_identifier_str oai:ri.conicet.gov.ar:11336/13119
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilaseSouza, Tatiana A. C. B.Okamoto, Débora N.Ruiz, Diego M.Oliveira, Lilian C. G.Kondo, Márcia Y.Tersario, Ivarne L. S.Juliano, Luizde Castro, Rosana EstherGouvea, Iuri E.Murakami, Mário T.Natrialba MagadiiExtracellular ProteaseHalophilismStructureStabilityKineticshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Nep (Natrialba magadii extracellular protease) is a halolysin-like peptidase secreted by the haloalkaliphilic archaeon N. magadii that exhibits optimal activity and stability in salt-saturated solutions. In this work, the effect of salt on the function and structure of Nep was investigated. In absence of salt, Nep became unfolded and aggregated, leading to the loss of activity. The enzyme did not recover its structural and functional properties even after restoring the ideal conditions for catalysis. At salt concentrations higher than 1 M (NaCl), Nep behaved as monomers in solution and its enzymatic activity displayed a nonlinear concave-up dependence with salt concentration resulting in a 20-fold activation at 4 M NaCl. Although transition from a high to a low-saline environment (3–1 M NaCl) did not affect its secondary structure contents, it diminished the enzyme stability and provoked large structural rearrangements, changing from an elongated shape at 3 M NaCl to a compact conformational state at 1 M NaCl. The thermodynamic analysis of peptide hydrolysis by Nep suggests a significant enzyme reorganization depending on the environmental salinity, which supports in solution SAXS and DLS studies. Moreover, solvent kinetic isotopic effect (SKIE) data indicates the general acid-base mechanism as the rate-limiting step for Nep catalysis, like classical serine-peptidases. All these data correlate the Nep conformational states with the enzymatic behavior providing a further understanding on the stability and structural determinants for the functioning of halolysins under different salinities.Fil: Souza, Tatiana A. C. B.. Centro Nacional de Pesquisas em Energia e Materiais; BrasilFil: Okamoto, Débora N.. Universidade de Sao Paulo; BrasilFil: Ruiz, Diego M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Oliveira, Lilian C. G.. Universidade de Sao Paulo; BrasilFil: Kondo, Márcia Y.. Universidade de Sao Paulo; BrasilFil: Tersario, Ivarne L. S.. Universidade de Mogi das Cruzes; BrasilFil: Juliano, Luiz. Universidade de Sao Paulo; BrasilFil: de Castro, Rosana Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Gouvea, Iuri E.. Universidade de Sao Paulo; BrasilFil: Murakami, Mário T.. Centro Nacional de Pesquisas em Energia e Materiais; BrasilElsevier Masson2012-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/13119Souza, Tatiana A. C. B.; Okamoto, Débora N.; Ruiz, Diego M.; Oliveira, Lilian C. G.; Kondo, Márcia Y.; et al.; Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase; Elsevier Masson; Biochimie; 94; 3; 3-2012; 798-8050300-9084enginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0300908411004500info:eu-repo/semantics/altIdentifier/doi/10.1016/j.biochi.2011.11.011info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:45:36Zoai:ri.conicet.gov.ar:11336/13119instacron: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 15:45:36.397CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
title Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
spellingShingle Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
Souza, Tatiana A. C. B.
Natrialba Magadii
Extracellular Protease
Halophilism
Structure
Stability
Kinetics
title_short Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
title_full Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
title_fullStr Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
title_full_unstemmed Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
title_sort Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase
dc.creator.none.fl_str_mv Souza, Tatiana A. C. B.
Okamoto, Débora N.
Ruiz, Diego M.
Oliveira, Lilian C. G.
Kondo, Márcia Y.
Tersario, Ivarne L. S.
Juliano, Luiz
de Castro, Rosana Esther
Gouvea, Iuri E.
Murakami, Mário T.
author Souza, Tatiana A. C. B.
author_facet Souza, Tatiana A. C. B.
Okamoto, Débora N.
Ruiz, Diego M.
Oliveira, Lilian C. G.
Kondo, Márcia Y.
Tersario, Ivarne L. S.
Juliano, Luiz
de Castro, Rosana Esther
Gouvea, Iuri E.
Murakami, Mário T.
author_role author
author2 Okamoto, Débora N.
Ruiz, Diego M.
Oliveira, Lilian C. G.
Kondo, Márcia Y.
Tersario, Ivarne L. S.
Juliano, Luiz
de Castro, Rosana Esther
Gouvea, Iuri E.
Murakami, Mário T.
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Natrialba Magadii
Extracellular Protease
Halophilism
Structure
Stability
Kinetics
topic Natrialba Magadii
Extracellular Protease
Halophilism
Structure
Stability
Kinetics
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Nep (Natrialba magadii extracellular protease) is a halolysin-like peptidase secreted by the haloalkaliphilic archaeon N. magadii that exhibits optimal activity and stability in salt-saturated solutions. In this work, the effect of salt on the function and structure of Nep was investigated. In absence of salt, Nep became unfolded and aggregated, leading to the loss of activity. The enzyme did not recover its structural and functional properties even after restoring the ideal conditions for catalysis. At salt concentrations higher than 1 M (NaCl), Nep behaved as monomers in solution and its enzymatic activity displayed a nonlinear concave-up dependence with salt concentration resulting in a 20-fold activation at 4 M NaCl. Although transition from a high to a low-saline environment (3–1 M NaCl) did not affect its secondary structure contents, it diminished the enzyme stability and provoked large structural rearrangements, changing from an elongated shape at 3 M NaCl to a compact conformational state at 1 M NaCl. The thermodynamic analysis of peptide hydrolysis by Nep suggests a significant enzyme reorganization depending on the environmental salinity, which supports in solution SAXS and DLS studies. Moreover, solvent kinetic isotopic effect (SKIE) data indicates the general acid-base mechanism as the rate-limiting step for Nep catalysis, like classical serine-peptidases. All these data correlate the Nep conformational states with the enzymatic behavior providing a further understanding on the stability and structural determinants for the functioning of halolysins under different salinities.
Fil: Souza, Tatiana A. C. B.. Centro Nacional de Pesquisas em Energia e Materiais; Brasil
Fil: Okamoto, Débora N.. Universidade de Sao Paulo; Brasil
Fil: Ruiz, Diego M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Oliveira, Lilian C. G.. Universidade de Sao Paulo; Brasil
Fil: Kondo, Márcia Y.. Universidade de Sao Paulo; Brasil
Fil: Tersario, Ivarne L. S.. Universidade de Mogi das Cruzes; Brasil
Fil: Juliano, Luiz. Universidade de Sao Paulo; Brasil
Fil: de Castro, Rosana Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Gouvea, Iuri E.. Universidade de Sao Paulo; Brasil
Fil: Murakami, Mário T.. Centro Nacional de Pesquisas em Energia e Materiais; Brasil
description Nep (Natrialba magadii extracellular protease) is a halolysin-like peptidase secreted by the haloalkaliphilic archaeon N. magadii that exhibits optimal activity and stability in salt-saturated solutions. In this work, the effect of salt on the function and structure of Nep was investigated. In absence of salt, Nep became unfolded and aggregated, leading to the loss of activity. The enzyme did not recover its structural and functional properties even after restoring the ideal conditions for catalysis. At salt concentrations higher than 1 M (NaCl), Nep behaved as monomers in solution and its enzymatic activity displayed a nonlinear concave-up dependence with salt concentration resulting in a 20-fold activation at 4 M NaCl. Although transition from a high to a low-saline environment (3–1 M NaCl) did not affect its secondary structure contents, it diminished the enzyme stability and provoked large structural rearrangements, changing from an elongated shape at 3 M NaCl to a compact conformational state at 1 M NaCl. The thermodynamic analysis of peptide hydrolysis by Nep suggests a significant enzyme reorganization depending on the environmental salinity, which supports in solution SAXS and DLS studies. Moreover, solvent kinetic isotopic effect (SKIE) data indicates the general acid-base mechanism as the rate-limiting step for Nep catalysis, like classical serine-peptidases. All these data correlate the Nep conformational states with the enzymatic behavior providing a further understanding on the stability and structural determinants for the functioning of halolysins under different salinities.
publishDate 2012
dc.date.none.fl_str_mv 2012-03
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/11336/13119
Souza, Tatiana A. C. B.; Okamoto, Débora N.; Ruiz, Diego M.; Oliveira, Lilian C. G.; Kondo, Márcia Y.; et al.; Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase; Elsevier Masson; Biochimie; 94; 3; 3-2012; 798-805
0300-9084
url http://hdl.handle.net/11336/13119
identifier_str_mv Souza, Tatiana A. C. B.; Okamoto, Débora N.; Ruiz, Diego M.; Oliveira, Lilian C. G.; Kondo, Márcia Y.; et al.; Correlation between catalysis and tertiary structure arrangement in an archaeal halophilic subtilase; Elsevier Masson; Biochimie; 94; 3; 3-2012; 798-805
0300-9084
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0300908411004500
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.biochi.2011.11.011
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 Elsevier Masson
publisher.none.fl_str_mv Elsevier Masson
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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