The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism

Autores
Infantes, Lourdes; Otero, Lisandro Horacio; Beassoni, Paola Rita; Boetsch, Cristhian; Lisa, Angela Teresita; Domenech, Carlos Eduardo; Albert, Armando
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen. It colonizes different tissues by the utilization of diverse mechanisms. One of these may involve the breakdown of the host cell membrane through the sequential action of haemolytic phospholipase-C and phosphorylcholine phosphatase (PchP). The action of haemolytic phospholipase-C on phosphatidylcholine produces phosphorylcholine, which is hydrolyzed to choline and inorganic phosphate by PchP. The available biochemical data on this enzyme demonstrate the involvement of two choline-binding sites in the catalytic cycle and in enzyme regulation. The crystal structure of P. aeruginosa PchP has been determined. It folds into three structural domains. The first domain harbors all the residues involved in catalysis and is well conserved among the haloacid dehalogenase (HAD) superfamily of proteins. The second domain is characteristic of PchP and is involved in the recognition of the choline moiety of the substrate. The third domain stabilizes the relative position of the other two. Fortuitously, the crystal structure of PchP captures molecules of Bis-Tris at the active site and at an additional site. This represents two catalytically relevant complexes with just one or two inhibitory Bis-Tris molecules and provides the basis of the PchP function and regulation. Site directed mutagenesis along with biochemical experiments corroborate the structural observations and demonstrate the interplay between different sites for choline recognition and inhibition. The structural comparison of PchP with other phosphatases of the HAD family provides a 3D-picture of the conserved catalytic cycle and the structural basis for the recognition of the diverse substrate molecules.
Fil: Infantes, Lourdes. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; España
Fil: Otero, Lisandro Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; España. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina
Fil: Beassoni, Paola Rita. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Boetsch, Cristhian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina
Fil: Lisa, Angela Teresita. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Domenech, Carlos Eduardo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Albert, Armando. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; España
Materia
amyloid
solid state
NMR
structure
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:11336/269278
Acceder
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network_acronym_str CONICETDig
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network_name_str CONICET Digital (CONICET)
spelling The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic MechanismInfantes, LourdesOtero, Lisandro HoracioBeassoni, Paola RitaBoetsch, CristhianLisa, Angela TeresitaDomenech, Carlos EduardoAlbert, Armandoamyloidsolid stateNMRstructurehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen. It colonizes different tissues by the utilization of diverse mechanisms. One of these may involve the breakdown of the host cell membrane through the sequential action of haemolytic phospholipase-C and phosphorylcholine phosphatase (PchP). The action of haemolytic phospholipase-C on phosphatidylcholine produces phosphorylcholine, which is hydrolyzed to choline and inorganic phosphate by PchP. The available biochemical data on this enzyme demonstrate the involvement of two choline-binding sites in the catalytic cycle and in enzyme regulation. The crystal structure of P. aeruginosa PchP has been determined. It folds into three structural domains. The first domain harbors all the residues involved in catalysis and is well conserved among the haloacid dehalogenase (HAD) superfamily of proteins. The second domain is characteristic of PchP and is involved in the recognition of the choline moiety of the substrate. The third domain stabilizes the relative position of the other two. Fortuitously, the crystal structure of PchP captures molecules of Bis-Tris at the active site and at an additional site. This represents two catalytically relevant complexes with just one or two inhibitory Bis-Tris molecules and provides the basis of the PchP function and regulation. Site directed mutagenesis along with biochemical experiments corroborate the structural observations and demonstrate the interplay between different sites for choline recognition and inhibition. The structural comparison of PchP with other phosphatases of the HAD family provides a 3D-picture of the conserved catalytic cycle and the structural basis for the recognition of the diverse substrate molecules.Fil: Infantes, Lourdes. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; EspañaFil: Otero, Lisandro Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; España. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Beassoni, Paola Rita. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Boetsch, Cristhian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; ArgentinaFil: Lisa, Angela Teresita. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Domenech, Carlos Eduardo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Albert, Armando. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; EspañaAcademic Press Ltd - Elsevier Science Ltd2012-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/269278Infantes, Lourdes; Otero, Lisandro Horacio; Beassoni, Paola Rita; Boetsch, Cristhian; Lisa, Angela Teresita; et al.; The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism; Academic Press Ltd - Elsevier Science Ltd; Journal of Molecular Biology; 423; 4; 11-2012; 503-5140022-2836CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0022283612006080?v=s5info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jmb.2012.07.024info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:19:24Zoai:ri.conicet.gov.ar:11336/269278instacron: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 15:19:25.263CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
title The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
spellingShingle The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
Infantes, Lourdes
amyloid
solid state
NMR
structure
title_short The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
title_full The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
title_fullStr The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
title_full_unstemmed The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
title_sort The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism
dc.creator.none.fl_str_mv Infantes, Lourdes
Otero, Lisandro Horacio
Beassoni, Paola Rita
Boetsch, Cristhian
Lisa, Angela Teresita
Domenech, Carlos Eduardo
Albert, Armando
author Infantes, Lourdes
author_facet Infantes, Lourdes
Otero, Lisandro Horacio
Beassoni, Paola Rita
Boetsch, Cristhian
Lisa, Angela Teresita
Domenech, Carlos Eduardo
Albert, Armando
author_role author
author2 Otero, Lisandro Horacio
Beassoni, Paola Rita
Boetsch, Cristhian
Lisa, Angela Teresita
Domenech, Carlos Eduardo
Albert, Armando
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv amyloid
solid state
NMR
structure
topic amyloid
solid state
NMR
structure
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen. It colonizes different tissues by the utilization of diverse mechanisms. One of these may involve the breakdown of the host cell membrane through the sequential action of haemolytic phospholipase-C and phosphorylcholine phosphatase (PchP). The action of haemolytic phospholipase-C on phosphatidylcholine produces phosphorylcholine, which is hydrolyzed to choline and inorganic phosphate by PchP. The available biochemical data on this enzyme demonstrate the involvement of two choline-binding sites in the catalytic cycle and in enzyme regulation. The crystal structure of P. aeruginosa PchP has been determined. It folds into three structural domains. The first domain harbors all the residues involved in catalysis and is well conserved among the haloacid dehalogenase (HAD) superfamily of proteins. The second domain is characteristic of PchP and is involved in the recognition of the choline moiety of the substrate. The third domain stabilizes the relative position of the other two. Fortuitously, the crystal structure of PchP captures molecules of Bis-Tris at the active site and at an additional site. This represents two catalytically relevant complexes with just one or two inhibitory Bis-Tris molecules and provides the basis of the PchP function and regulation. Site directed mutagenesis along with biochemical experiments corroborate the structural observations and demonstrate the interplay between different sites for choline recognition and inhibition. The structural comparison of PchP with other phosphatases of the HAD family provides a 3D-picture of the conserved catalytic cycle and the structural basis for the recognition of the diverse substrate molecules.
Fil: Infantes, Lourdes. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; España
Fil: Otero, Lisandro Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; España. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina
Fil: Beassoni, Paola Rita. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Boetsch, Cristhian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina
Fil: Lisa, Angela Teresita. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Domenech, Carlos Eduardo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Albert, Armando. Consejo Superior de Investigaciones Científicas. Instituto de Química Física "Rocasolano"; España
description Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen. It colonizes different tissues by the utilization of diverse mechanisms. One of these may involve the breakdown of the host cell membrane through the sequential action of haemolytic phospholipase-C and phosphorylcholine phosphatase (PchP). The action of haemolytic phospholipase-C on phosphatidylcholine produces phosphorylcholine, which is hydrolyzed to choline and inorganic phosphate by PchP. The available biochemical data on this enzyme demonstrate the involvement of two choline-binding sites in the catalytic cycle and in enzyme regulation. The crystal structure of P. aeruginosa PchP has been determined. It folds into three structural domains. The first domain harbors all the residues involved in catalysis and is well conserved among the haloacid dehalogenase (HAD) superfamily of proteins. The second domain is characteristic of PchP and is involved in the recognition of the choline moiety of the substrate. The third domain stabilizes the relative position of the other two. Fortuitously, the crystal structure of PchP captures molecules of Bis-Tris at the active site and at an additional site. This represents two catalytically relevant complexes with just one or two inhibitory Bis-Tris molecules and provides the basis of the PchP function and regulation. Site directed mutagenesis along with biochemical experiments corroborate the structural observations and demonstrate the interplay between different sites for choline recognition and inhibition. The structural comparison of PchP with other phosphatases of the HAD family provides a 3D-picture of the conserved catalytic cycle and the structural basis for the recognition of the diverse substrate molecules.
publishDate 2012
dc.date.none.fl_str_mv 2012-11
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/11336/269278
Infantes, Lourdes; Otero, Lisandro Horacio; Beassoni, Paola Rita; Boetsch, Cristhian; Lisa, Angela Teresita; et al.; The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism; Academic Press Ltd - Elsevier Science Ltd; Journal of Molecular Biology; 423; 4; 11-2012; 503-514
0022-2836
CONICET Digital
CONICET
url http://hdl.handle.net/11336/269278
identifier_str_mv Infantes, Lourdes; Otero, Lisandro Horacio; Beassoni, Paola Rita; Boetsch, Cristhian; Lisa, Angela Teresita; et al.; The Structural Domains of Pseudomonas aeruginosa Phosphorylcholine Phosphatase Cooperate in Substrate Hydrolysis: 3D Structure and Enzymatic Mechanism; Academic Press Ltd - Elsevier Science Ltd; Journal of Molecular Biology; 423; 4; 11-2012; 503-514
0022-2836
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://www.sciencedirect.com/science/article/pii/S0022283612006080?v=s5
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jmb.2012.07.024
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
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dc.publisher.none.fl_str_mv Academic Press Ltd - Elsevier Science Ltd
publisher.none.fl_str_mv Academic Press Ltd - Elsevier Science Ltd
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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