Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex

Autores
Trejo, Sebastián Alejandro; López, Laura María Isabel; Caffini, Néstor Oscar; Natalucci, Claudia Luisa; Canals, Francesc; Avilés, Francesc X.
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Asclepain f is a papain-like protease previously isolated and characterized from latex of Asclepias fruticosa. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZα vector and expressed in Pichia pastoris. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.
Centro de Investigación de Proteínas Vegetales
Materia
Ciencias Exactas
Biología
Asclepias fruticosa
Gomphocarpus fruticosus subsp. fruticosus
Plant latex
Cysteine endopeptidase
Cloning
Overexpression in Pichia pastoris
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/142367
Acceder
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oai_identifier_str oai:sedici.unlp.edu.ar:10915/142367
network_acronym_str SEDICI
repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latexTrejo, Sebastián AlejandroLópez, Laura María IsabelCaffini, Néstor OscarNatalucci, Claudia LuisaCanals, FrancescAvilés, Francesc X.Ciencias ExactasBiologíaAsclepias fruticosaGomphocarpus fruticosus subsp. fruticosusPlant latexCysteine endopeptidaseCloningOverexpression in Pichia pastorisAsclepain f is a papain-like protease previously isolated and characterized from latex of <i>Asclepias fruticosa</i>. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZα vector and expressed in <i>Pichia pastoris</i>. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.Centro de Investigación de Proteínas Vegetales2009-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf319-328http://sedici.unlp.edu.ar/handle/10915/142367enginfo:eu-repo/semantics/altIdentifier/issn/1432-2048info:eu-repo/semantics/altIdentifier/issn/0032-0935info:eu-repo/semantics/altIdentifier/doi/10.1007/s00425-009-0942-2info:eu-repo/semantics/altIdentifier/pmid/19455353info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International (CC BY 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:32:32Zoai:sedici.unlp.edu.ar:10915/142367Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:32:33.073SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
title Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
spellingShingle Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
Trejo, Sebastián Alejandro
Ciencias Exactas
Biología
Asclepias fruticosa
Gomphocarpus fruticosus subsp. fruticosus
Plant latex
Cysteine endopeptidase
Cloning
Overexpression in Pichia pastoris
title_short Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
title_full Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
title_fullStr Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
title_full_unstemmed Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
title_sort Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
dc.creator.none.fl_str_mv Trejo, Sebastián Alejandro
López, Laura María Isabel
Caffini, Néstor Oscar
Natalucci, Claudia Luisa
Canals, Francesc
Avilés, Francesc X.
author Trejo, Sebastián Alejandro
author_facet Trejo, Sebastián Alejandro
López, Laura María Isabel
Caffini, Néstor Oscar
Natalucci, Claudia Luisa
Canals, Francesc
Avilés, Francesc X.
author_role author
author2 López, Laura María Isabel
Caffini, Néstor Oscar
Natalucci, Claudia Luisa
Canals, Francesc
Avilés, Francesc X.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Ciencias Exactas
Biología
Asclepias fruticosa
Gomphocarpus fruticosus subsp. fruticosus
Plant latex
Cysteine endopeptidase
Cloning
Overexpression in Pichia pastoris
topic Ciencias Exactas
Biología
Asclepias fruticosa
Gomphocarpus fruticosus subsp. fruticosus
Plant latex
Cysteine endopeptidase
Cloning
Overexpression in Pichia pastoris
dc.description.none.fl_txt_mv Asclepain f is a papain-like protease previously isolated and characterized from latex of <i>Asclepias fruticosa</i>. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZα vector and expressed in <i>Pichia pastoris</i>. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.
Centro de Investigación de Proteínas Vegetales
description Asclepain f is a papain-like protease previously isolated and characterized from latex of <i>Asclepias fruticosa</i>. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZα vector and expressed in <i>Pichia pastoris</i>. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.
publishDate 2009
dc.date.none.fl_str_mv 2009-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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://sedici.unlp.edu.ar/handle/10915/142367
url http://sedici.unlp.edu.ar/handle/10915/142367
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/1432-2048
info:eu-repo/semantics/altIdentifier/issn/0032-0935
info:eu-repo/semantics/altIdentifier/doi/10.1007/s00425-009-0942-2
info:eu-repo/semantics/altIdentifier/pmid/19455353
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International (CC BY 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International (CC BY 4.0)
dc.format.none.fl_str_mv application/pdf
319-328
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
reponame_str SEDICI (UNLP)
collection SEDICI (UNLP)
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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