Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor

Autores
Covaleda Cortes, Giovanni; Hernández, Martha; Trejo, Sebastian Alejandro; Mansur, Manuel; Rodriguez Calado, Sergi; Garcia Pardo, Javier; Lorenzo, Julia; Vendrell, Josep; Chávez, María Ángeles; Alonso Del Rivero, Maday; Avilés, Francesc Xavier
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A very powerful proteinaceous inhibitor of metallocarboxypeptidases has been isolated from the marine snail Nerita versicolor and characterized in depth. The most abundant of four, very similar isoforms, NvCla, was taken as reference and N-terminally sequenced to obtain a 372-nucleotide band coding for the protein cDNA. The mature protein contains 53 residues and three disulphide bonds. NvCIa and the other isoforms show an exceptionally high inhibitory capacity of around 1.8 pM for human Carboxypeptidase A1 (hCPA1) and for other A-like members of the M14 CPAsubfamily, whereas a twofold decrease in inhibitory potency is observed for carboxypeptidase B-like members as hCPB and hTAFIa. A recombinant form, rNvCI, was produced in high yield and HPLC, mass spectrometry and spectroscopic analyses by CD and NMR indicated its homogeneous, compact and thermally resistant nature. Using antibodies raised with rNvCI and histochemical analyses, a preferential distribution of the inhibitor in the surface regions of the animal body was observed, particularly nearby the open entrance of the shell and gut, suggesting its involvement in biological defense mechanisms. The properties of this strong, small and stable inhibitor of metallocarboxypeptidases envisage potentialities for its direct applicability, as well as leading or minimized forms, in biotechnological/biomedical uses.
Fil: Covaleda Cortes, Giovanni. Universitat Autònoma de Barcelona; España
Fil: Hernández, Martha. Universidad de Concepción; Chile
Fil: Trejo, Sebastian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universitat Autònoma de Barcelona; España. YPF - Tecnología; Argentina
Fil: Mansur, Manuel. Universitat Autònoma de Barcelona; España
Fil: Rodriguez Calado, Sergi. Universitat Autònoma de Barcelona; España
Fil: Garcia Pardo, Javier. Universitat Autònoma de Barcelona; España
Fil: Lorenzo, Julia. Universitat Autònoma de Barcelona; España
Fil: Vendrell, Josep. Universitat Autònoma de Barcelona; España
Fil: Chávez, María Ángeles. Universidad de la Habana. Facultad de Biología. Centro de Estudios de Las Proteinas; Cuba
Fil: Alonso Del Rivero, Maday. Universidad de la Habana. Facultad de Biología. Centro de Estudios de Las Proteinas; Cuba
Fil: Avilés, Francesc Xavier. Universitat Autònoma de Barcelona; España
Materia
BIOMEDICAL APPLICATIONS
BIOTECHNOLOGICAL
CARBOXYPEPTIDASE
NERITA VERSICOLOR
PICOMOLAR INHIBITION
PROTEINACEOUS INHIBITOR
RECOMBINANT PRODUCTION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/159977
Acceder
id CONICETDig_3a74b5de42cf9db7680bc3e56762d733
oai_identifier_str oai:ri.conicet.gov.ar:11336/159977
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolorCovaleda Cortes, GiovanniHernández, MarthaTrejo, Sebastian AlejandroMansur, ManuelRodriguez Calado, SergiGarcia Pardo, JavierLorenzo, JuliaVendrell, JosepChávez, María ÁngelesAlonso Del Rivero, MadayAvilés, Francesc XavierBIOMEDICAL APPLICATIONSBIOTECHNOLOGICALCARBOXYPEPTIDASENERITA VERSICOLORPICOMOLAR INHIBITIONPROTEINACEOUS INHIBITORRECOMBINANT PRODUCTIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1A very powerful proteinaceous inhibitor of metallocarboxypeptidases has been isolated from the marine snail Nerita versicolor and characterized in depth. The most abundant of four, very similar isoforms, NvCla, was taken as reference and N-terminally sequenced to obtain a 372-nucleotide band coding for the protein cDNA. The mature protein contains 53 residues and three disulphide bonds. NvCIa and the other isoforms show an exceptionally high inhibitory capacity of around 1.8 pM for human Carboxypeptidase A1 (hCPA1) and for other A-like members of the M14 CPAsubfamily, whereas a twofold decrease in inhibitory potency is observed for carboxypeptidase B-like members as hCPB and hTAFIa. A recombinant form, rNvCI, was produced in high yield and HPLC, mass spectrometry and spectroscopic analyses by CD and NMR indicated its homogeneous, compact and thermally resistant nature. Using antibodies raised with rNvCI and histochemical analyses, a preferential distribution of the inhibitor in the surface regions of the animal body was observed, particularly nearby the open entrance of the shell and gut, suggesting its involvement in biological defense mechanisms. The properties of this strong, small and stable inhibitor of metallocarboxypeptidases envisage potentialities for its direct applicability, as well as leading or minimized forms, in biotechnological/biomedical uses.Fil: Covaleda Cortes, Giovanni. Universitat Autònoma de Barcelona; EspañaFil: Hernández, Martha. Universidad de Concepción; ChileFil: Trejo, Sebastian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universitat Autònoma de Barcelona; España. YPF - Tecnología; ArgentinaFil: Mansur, Manuel. Universitat Autònoma de Barcelona; EspañaFil: Rodriguez Calado, Sergi. Universitat Autònoma de Barcelona; EspañaFil: Garcia Pardo, Javier. Universitat Autònoma de Barcelona; EspañaFil: Lorenzo, Julia. Universitat Autònoma de Barcelona; EspañaFil: Vendrell, Josep. Universitat Autònoma de Barcelona; EspañaFil: Chávez, María Ángeles. Universidad de la Habana. Facultad de Biología. Centro de Estudios de Las Proteinas; CubaFil: Alonso Del Rivero, Maday. Universidad de la Habana. Facultad de Biología. Centro de Estudios de Las Proteinas; CubaFil: Avilés, Francesc Xavier. Universitat Autònoma de Barcelona; EspañaMultidisciplinary Digital Publishing Institute2019-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/159977Covaleda Cortes, Giovanni; Hernández, Martha; Trejo, Sebastian Alejandro; Mansur, Manuel; Rodriguez Calado, Sergi; et al.; Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor; Multidisciplinary Digital Publishing Institute; Marine Drugs; 17; 9; 8-2019; 1-191660-3397CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/1660-3397/17/9/511info:eu-repo/semantics/altIdentifier/doi/10.3390/md17090511info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:15:47Zoai:ri.conicet.gov.ar:11336/159977instacron: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:34982025-10-22 11:15:48.238CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
title Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
spellingShingle Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
Covaleda Cortes, Giovanni
BIOMEDICAL APPLICATIONS
BIOTECHNOLOGICAL
CARBOXYPEPTIDASE
NERITA VERSICOLOR
PICOMOLAR INHIBITION
PROTEINACEOUS INHIBITOR
RECOMBINANT PRODUCTION
title_short Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
title_full Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
title_fullStr Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
title_full_unstemmed Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
title_sort Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor
dc.creator.none.fl_str_mv Covaleda Cortes, Giovanni
Hernández, Martha
Trejo, Sebastian Alejandro
Mansur, Manuel
Rodriguez Calado, Sergi
Garcia Pardo, Javier
Lorenzo, Julia
Vendrell, Josep
Chávez, María Ángeles
Alonso Del Rivero, Maday
Avilés, Francesc Xavier
author Covaleda Cortes, Giovanni
author_facet Covaleda Cortes, Giovanni
Hernández, Martha
Trejo, Sebastian Alejandro
Mansur, Manuel
Rodriguez Calado, Sergi
Garcia Pardo, Javier
Lorenzo, Julia
Vendrell, Josep
Chávez, María Ángeles
Alonso Del Rivero, Maday
Avilés, Francesc Xavier
author_role author
author2 Hernández, Martha
Trejo, Sebastian Alejandro
Mansur, Manuel
Rodriguez Calado, Sergi
Garcia Pardo, Javier
Lorenzo, Julia
Vendrell, Josep
Chávez, María Ángeles
Alonso Del Rivero, Maday
Avilés, Francesc Xavier
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv BIOMEDICAL APPLICATIONS
BIOTECHNOLOGICAL
CARBOXYPEPTIDASE
NERITA VERSICOLOR
PICOMOLAR INHIBITION
PROTEINACEOUS INHIBITOR
RECOMBINANT PRODUCTION
topic BIOMEDICAL APPLICATIONS
BIOTECHNOLOGICAL
CARBOXYPEPTIDASE
NERITA VERSICOLOR
PICOMOLAR INHIBITION
PROTEINACEOUS INHIBITOR
RECOMBINANT PRODUCTION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv A very powerful proteinaceous inhibitor of metallocarboxypeptidases has been isolated from the marine snail Nerita versicolor and characterized in depth. The most abundant of four, very similar isoforms, NvCla, was taken as reference and N-terminally sequenced to obtain a 372-nucleotide band coding for the protein cDNA. The mature protein contains 53 residues and three disulphide bonds. NvCIa and the other isoforms show an exceptionally high inhibitory capacity of around 1.8 pM for human Carboxypeptidase A1 (hCPA1) and for other A-like members of the M14 CPAsubfamily, whereas a twofold decrease in inhibitory potency is observed for carboxypeptidase B-like members as hCPB and hTAFIa. A recombinant form, rNvCI, was produced in high yield and HPLC, mass spectrometry and spectroscopic analyses by CD and NMR indicated its homogeneous, compact and thermally resistant nature. Using antibodies raised with rNvCI and histochemical analyses, a preferential distribution of the inhibitor in the surface regions of the animal body was observed, particularly nearby the open entrance of the shell and gut, suggesting its involvement in biological defense mechanisms. The properties of this strong, small and stable inhibitor of metallocarboxypeptidases envisage potentialities for its direct applicability, as well as leading or minimized forms, in biotechnological/biomedical uses.
Fil: Covaleda Cortes, Giovanni. Universitat Autònoma de Barcelona; España
Fil: Hernández, Martha. Universidad de Concepción; Chile
Fil: Trejo, Sebastian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universitat Autònoma de Barcelona; España. YPF - Tecnología; Argentina
Fil: Mansur, Manuel. Universitat Autònoma de Barcelona; España
Fil: Rodriguez Calado, Sergi. Universitat Autònoma de Barcelona; España
Fil: Garcia Pardo, Javier. Universitat Autònoma de Barcelona; España
Fil: Lorenzo, Julia. Universitat Autònoma de Barcelona; España
Fil: Vendrell, Josep. Universitat Autònoma de Barcelona; España
Fil: Chávez, María Ángeles. Universidad de la Habana. Facultad de Biología. Centro de Estudios de Las Proteinas; Cuba
Fil: Alonso Del Rivero, Maday. Universidad de la Habana. Facultad de Biología. Centro de Estudios de Las Proteinas; Cuba
Fil: Avilés, Francesc Xavier. Universitat Autònoma de Barcelona; España
description A very powerful proteinaceous inhibitor of metallocarboxypeptidases has been isolated from the marine snail Nerita versicolor and characterized in depth. The most abundant of four, very similar isoforms, NvCla, was taken as reference and N-terminally sequenced to obtain a 372-nucleotide band coding for the protein cDNA. The mature protein contains 53 residues and three disulphide bonds. NvCIa and the other isoforms show an exceptionally high inhibitory capacity of around 1.8 pM for human Carboxypeptidase A1 (hCPA1) and for other A-like members of the M14 CPAsubfamily, whereas a twofold decrease in inhibitory potency is observed for carboxypeptidase B-like members as hCPB and hTAFIa. A recombinant form, rNvCI, was produced in high yield and HPLC, mass spectrometry and spectroscopic analyses by CD and NMR indicated its homogeneous, compact and thermally resistant nature. Using antibodies raised with rNvCI and histochemical analyses, a preferential distribution of the inhibitor in the surface regions of the animal body was observed, particularly nearby the open entrance of the shell and gut, suggesting its involvement in biological defense mechanisms. The properties of this strong, small and stable inhibitor of metallocarboxypeptidases envisage potentialities for its direct applicability, as well as leading or minimized forms, in biotechnological/biomedical uses.
publishDate 2019
dc.date.none.fl_str_mv 2019-08
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/11336/159977
Covaleda Cortes, Giovanni; Hernández, Martha; Trejo, Sebastian Alejandro; Mansur, Manuel; Rodriguez Calado, Sergi; et al.; Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor; Multidisciplinary Digital Publishing Institute; Marine Drugs; 17; 9; 8-2019; 1-19
1660-3397
CONICET Digital
CONICET
url http://hdl.handle.net/11336/159977
identifier_str_mv Covaleda Cortes, Giovanni; Hernández, Martha; Trejo, Sebastian Alejandro; Mansur, Manuel; Rodriguez Calado, Sergi; et al.; Characterization, recombinant production and structure-function analysis of NvCI, a picomolar metallocarboxypeptidase inhibitor from the marine nnail Nerita versicolor; Multidisciplinary Digital Publishing Institute; Marine Drugs; 17; 9; 8-2019; 1-19
1660-3397
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.mdpi.com/1660-3397/17/9/511
info:eu-repo/semantics/altIdentifier/doi/10.3390/md17090511
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
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 13.229304

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