High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase

Autores
Dematteis, Massimiliano; Bovi, Michele; Bertinelli, Mattia; Bertini, Edoardo; Destefanis, Laura; Carrizo Garcia, Maria Elena; Capaldi, Stefano; Monaco, Hugo
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Lipocalin-type prostaglandin D synthase (L-PGDS) catalyzes the isomerization of the 9,11-endoperoxide group of PGH2 (prostaglandin H2) to produce PGD2 (prostaglandin D2) with 9-hydroxy and 11-keto groups. The product of the reaction, PGD2, is the precursor of several metabolites involved in many regulatory events. L-PGDS, the first member of the important lipocalin family to be recognized as an enzyme, is also able to bind and transport small hydrophobic molecules and was formerly known as [beta]-trace protein, the second most abundant protein in human cerebrospinal fluid. Previous structural work on the mouse and human proteins has focused on the identification of the amino acids responsible and the proposal of a mechanism for catalysis. In this paper, the X-ray structures of the apo and holo forms (bound to PEG) of the C65A mutant of human L-PGDS at 1.40 Å resolution and of the double mutant C65A/K59A at 1.60 Å resolution are reported. The apo forms of the double mutants C65A/W54F and C65A/W112F and the triple mutant C65A/W54F/W112F have also been studied. Mutation of the lysine residue does not seem to affect the binding of PEG to the ligand-binding cavity, and mutation of a single or both tryptophans appears to have the same effect on the position of these two aromatic residues at the entrance to the cavity. A solvent molecule has also been identified in an invariant position in the cavity of virtually all of the molecules present in the nine asymmetric units of the crystals that have been examined. Taken together, these observations indicate that the residues that have been mutated indeed appear to play a role in the entrance-exit process of the substrate and/or other ligands into/out of the binding cavity of the lipocalin.
Fil: Dematteis, Massimiliano. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Bovi, Michele. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Bertinelli, Mattia. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Bertini, Edoardo. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Destefanis, Laura. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Carrizo Garcia, Maria Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Capaldi, Stefano. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Monaco, Hugo. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Materia
Lipocalin-Type Prostaglandin D Synthase
Mutants
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:11336/32094
Acceder
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network_acronym_str CONICETDig
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network_name_str CONICET Digital (CONICET)
spelling High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthaseDematteis, MassimilianoBovi, MicheleBertinelli, MattiaBertini, EdoardoDestefanis, LauraCarrizo Garcia, Maria ElenaCapaldi, StefanoMonaco, HugoLipocalin-Type Prostaglandin D SynthaseMutantshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Lipocalin-type prostaglandin D synthase (L-PGDS) catalyzes the isomerization of the 9,11-endoperoxide group of PGH2 (prostaglandin H2) to produce PGD2 (prostaglandin D2) with 9-hydroxy and 11-keto groups. The product of the reaction, PGD2, is the precursor of several metabolites involved in many regulatory events. L-PGDS, the first member of the important lipocalin family to be recognized as an enzyme, is also able to bind and transport small hydrophobic molecules and was formerly known as [beta]-trace protein, the second most abundant protein in human cerebrospinal fluid. Previous structural work on the mouse and human proteins has focused on the identification of the amino acids responsible and the proposal of a mechanism for catalysis. In this paper, the X-ray structures of the apo and holo forms (bound to PEG) of the C65A mutant of human L-PGDS at 1.40 Å resolution and of the double mutant C65A/K59A at 1.60 Å resolution are reported. The apo forms of the double mutants C65A/W54F and C65A/W112F and the triple mutant C65A/W54F/W112F have also been studied. Mutation of the lysine residue does not seem to affect the binding of PEG to the ligand-binding cavity, and mutation of a single or both tryptophans appears to have the same effect on the position of these two aromatic residues at the entrance to the cavity. A solvent molecule has also been identified in an invariant position in the cavity of virtually all of the molecules present in the nine asymmetric units of the crystals that have been examined. Taken together, these observations indicate that the residues that have been mutated indeed appear to play a role in the entrance-exit process of the substrate and/or other ligands into/out of the binding cavity of the lipocalin.Fil: Dematteis, Massimiliano. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; ItaliaFil: Bovi, Michele. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; ItaliaFil: Bertinelli, Mattia. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; ItaliaFil: Bertini, Edoardo. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; ItaliaFil: Destefanis, Laura. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; ItaliaFil: Carrizo Garcia, Maria Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Capaldi, Stefano. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; ItaliaFil: Monaco, Hugo. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; ItaliaWiley Blackwell Publishing, Inc2014-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/32094Monaco, Hugo; Capaldi, Stefano; Carrizo Garcia, Maria Elena; Destefanis, Laura; Bertini, Edoardo; Bertinelli, Mattia; et al.; High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase; Wiley Blackwell Publishing, Inc; Acta Crystallographica Section D-biological Crystallography; 70; 5-2014; 2125-21380907-4449CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1107/S1399004714012462info:eu-repo/semantics/altIdentifier/url/http://scripts.iucr.org/cgi-bin/paper?S1399004714012462info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:47:14Zoai:ri.conicet.gov.ar:11336/32094instacron: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 09:47:14.467CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
title High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
spellingShingle High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
Dematteis, Massimiliano
Lipocalin-Type Prostaglandin D Synthase
Mutants
title_short High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
title_full High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
title_fullStr High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
title_full_unstemmed High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
title_sort High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase
dc.creator.none.fl_str_mv Dematteis, Massimiliano
Bovi, Michele
Bertinelli, Mattia
Bertini, Edoardo
Destefanis, Laura
Carrizo Garcia, Maria Elena
Capaldi, Stefano
Monaco, Hugo
author Dematteis, Massimiliano
author_facet Dematteis, Massimiliano
Bovi, Michele
Bertinelli, Mattia
Bertini, Edoardo
Destefanis, Laura
Carrizo Garcia, Maria Elena
Capaldi, Stefano
Monaco, Hugo
author_role author
author2 Bovi, Michele
Bertinelli, Mattia
Bertini, Edoardo
Destefanis, Laura
Carrizo Garcia, Maria Elena
Capaldi, Stefano
Monaco, Hugo
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Lipocalin-Type Prostaglandin D Synthase
Mutants
topic Lipocalin-Type Prostaglandin D Synthase
Mutants
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Lipocalin-type prostaglandin D synthase (L-PGDS) catalyzes the isomerization of the 9,11-endoperoxide group of PGH2 (prostaglandin H2) to produce PGD2 (prostaglandin D2) with 9-hydroxy and 11-keto groups. The product of the reaction, PGD2, is the precursor of several metabolites involved in many regulatory events. L-PGDS, the first member of the important lipocalin family to be recognized as an enzyme, is also able to bind and transport small hydrophobic molecules and was formerly known as [beta]-trace protein, the second most abundant protein in human cerebrospinal fluid. Previous structural work on the mouse and human proteins has focused on the identification of the amino acids responsible and the proposal of a mechanism for catalysis. In this paper, the X-ray structures of the apo and holo forms (bound to PEG) of the C65A mutant of human L-PGDS at 1.40 Å resolution and of the double mutant C65A/K59A at 1.60 Å resolution are reported. The apo forms of the double mutants C65A/W54F and C65A/W112F and the triple mutant C65A/W54F/W112F have also been studied. Mutation of the lysine residue does not seem to affect the binding of PEG to the ligand-binding cavity, and mutation of a single or both tryptophans appears to have the same effect on the position of these two aromatic residues at the entrance to the cavity. A solvent molecule has also been identified in an invariant position in the cavity of virtually all of the molecules present in the nine asymmetric units of the crystals that have been examined. Taken together, these observations indicate that the residues that have been mutated indeed appear to play a role in the entrance-exit process of the substrate and/or other ligands into/out of the binding cavity of the lipocalin.
Fil: Dematteis, Massimiliano. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Bovi, Michele. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Bertinelli, Mattia. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Bertini, Edoardo. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Destefanis, Laura. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Carrizo Garcia, Maria Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Capaldi, Stefano. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
Fil: Monaco, Hugo. Universita Di Verona. Dipartimento Scientífico E Tecnológico. Laboratorio Di Biocristallografia; Italia
description Lipocalin-type prostaglandin D synthase (L-PGDS) catalyzes the isomerization of the 9,11-endoperoxide group of PGH2 (prostaglandin H2) to produce PGD2 (prostaglandin D2) with 9-hydroxy and 11-keto groups. The product of the reaction, PGD2, is the precursor of several metabolites involved in many regulatory events. L-PGDS, the first member of the important lipocalin family to be recognized as an enzyme, is also able to bind and transport small hydrophobic molecules and was formerly known as [beta]-trace protein, the second most abundant protein in human cerebrospinal fluid. Previous structural work on the mouse and human proteins has focused on the identification of the amino acids responsible and the proposal of a mechanism for catalysis. In this paper, the X-ray structures of the apo and holo forms (bound to PEG) of the C65A mutant of human L-PGDS at 1.40 Å resolution and of the double mutant C65A/K59A at 1.60 Å resolution are reported. The apo forms of the double mutants C65A/W54F and C65A/W112F and the triple mutant C65A/W54F/W112F have also been studied. Mutation of the lysine residue does not seem to affect the binding of PEG to the ligand-binding cavity, and mutation of a single or both tryptophans appears to have the same effect on the position of these two aromatic residues at the entrance to the cavity. A solvent molecule has also been identified in an invariant position in the cavity of virtually all of the molecules present in the nine asymmetric units of the crystals that have been examined. Taken together, these observations indicate that the residues that have been mutated indeed appear to play a role in the entrance-exit process of the substrate and/or other ligands into/out of the binding cavity of the lipocalin.
publishDate 2014
dc.date.none.fl_str_mv 2014-05
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/11336/32094
Monaco, Hugo; Capaldi, Stefano; Carrizo Garcia, Maria Elena; Destefanis, Laura; Bertini, Edoardo; Bertinelli, Mattia; et al.; High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase; Wiley Blackwell Publishing, Inc; Acta Crystallographica Section D-biological Crystallography; 70; 5-2014; 2125-2138
0907-4449
CONICET Digital
CONICET
url http://hdl.handle.net/11336/32094
identifier_str_mv Monaco, Hugo; Capaldi, Stefano; Carrizo Garcia, Maria Elena; Destefanis, Laura; Bertini, Edoardo; Bertinelli, Mattia; et al.; High-resolution structures of mutants of residues that affectaccess to the ligand-binding cavity of human lipocalin-typeprostaglandin D synthase; Wiley Blackwell Publishing, Inc; Acta Crystallographica Section D-biological Crystallography; 70; 5-2014; 2125-2138
0907-4449
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1107/S1399004714012462
info:eu-repo/semantics/altIdentifier/url/http://scripts.iucr.org/cgi-bin/paper?S1399004714012462
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 Wiley Blackwell Publishing, Inc
publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
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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