Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez

Autores
Bruno, Mariela Anahí; Trejo, Sebastian Alejandro; Aviles, Francesc Xavier; Caffini, Nestor Oscar; Lopez, Laura Maria Isabel
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Fruits of Bromelia hieronymi, a tropical South American plant, possess a high content of peptidases with potential biotechnological uses. Total RNA was extracted from unripe fruits and peptidase cDNA was obtained by 3′RACE-PCR. The consensus sequence of the cysteine peptidase cDNA contained 875 bp, the 690 first ones codifying for a hypothetical polypeptide chain of the mature peptidase, named Bh-CP1 (molecular mass 24.773 kDa, pI 8.6, extinction molar coefficient 58,705 M−1 cm−1). Bh-CP1 sequence shows a high percentage of identity with those of other cysteine plant proteases. The presence of highly preserved residues is observed, like those forming the catalytic site (Gln19, Cys25, His159, and Asn175, papain numbering), as well as other six Cys residues, involved in the formation of disulfide bounds. Molecular modeling results suggest the enzyme belongs to the α + β class of proteins, with two disulfide bridges (Cys23–Cys63 and Cys57–Cys96) in the α domain, while the β domain is stabilized by another disulfide bridge (Cys153–Cys203). Additionally, peptide mass fingerprints (PMFs) of the three peptidases previously isolated from B. hieronymi fruits (namely hieronymain I, II, and III) were performed and compared with the theoretical fingerprint of PMF of Bh-CP1, showing a partial matching between the in silico-translated protein and hieronymain II.
Fil: Bruno, Mariela Anahí. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Trejo, Sebastian Alejandro. Universidad Autonoma de Barcelona. Departamento de Biología; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Aviles, Francesc Xavier. Universidad Autonoma de Barcelona. Departamento de Biología; España
Fil: Caffini, Nestor Oscar. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina
Fil: Lopez, Laura Maria Isabel. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
BROMELIA HIERONYMI
BROMELIACEAE
CLONING
CYSTEINE ENDOPEPTIDASE
HIERONYMAIN II
PROTEOMIC TOOLS
SEQUENCING
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:11336/95875
Acceder
id CONICETDig_90418a6db709317918ca4607c28c1f0a
oai_identifier_str oai:ri.conicet.gov.ar:11336/95875
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mezBruno, Mariela AnahíTrejo, Sebastian AlejandroAviles, Francesc XavierCaffini, Nestor OscarLopez, Laura Maria IsabelBROMELIA HIERONYMIBROMELIACEAECLONINGCYSTEINE ENDOPEPTIDASEHIERONYMAIN IIPROTEOMIC TOOLSSEQUENCINGhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Fruits of Bromelia hieronymi, a tropical South American plant, possess a high content of peptidases with potential biotechnological uses. Total RNA was extracted from unripe fruits and peptidase cDNA was obtained by 3′RACE-PCR. The consensus sequence of the cysteine peptidase cDNA contained 875 bp, the 690 first ones codifying for a hypothetical polypeptide chain of the mature peptidase, named Bh-CP1 (molecular mass 24.773 kDa, pI 8.6, extinction molar coefficient 58,705 M−1 cm−1). Bh-CP1 sequence shows a high percentage of identity with those of other cysteine plant proteases. The presence of highly preserved residues is observed, like those forming the catalytic site (Gln19, Cys25, His159, and Asn175, papain numbering), as well as other six Cys residues, involved in the formation of disulfide bounds. Molecular modeling results suggest the enzyme belongs to the α + β class of proteins, with two disulfide bridges (Cys23–Cys63 and Cys57–Cys96) in the α domain, while the β domain is stabilized by another disulfide bridge (Cys153–Cys203). Additionally, peptide mass fingerprints (PMFs) of the three peptidases previously isolated from B. hieronymi fruits (namely hieronymain I, II, and III) were performed and compared with the theoretical fingerprint of PMF of Bh-CP1, showing a partial matching between the in silico-translated protein and hieronymain II.Fil: Bruno, Mariela Anahí. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Trejo, Sebastian Alejandro. Universidad Autonoma de Barcelona. Departamento de Biología; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Aviles, Francesc Xavier. Universidad Autonoma de Barcelona. Departamento de Biología; EspañaFil: Caffini, Nestor Oscar. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; ArgentinaFil: Lopez, Laura Maria Isabel. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaHumana Press2011-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/95875Bruno, Mariela Anahí; Trejo, Sebastian Alejandro; Aviles, Francesc Xavier; Caffini, Nestor Oscar; Lopez, Laura Maria Isabel; Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez; Humana Press; Applied Biochemistry And Biotechnology; 165; 2; 9-2011; 583-5930273-2289CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007/s12010-011-9277-0info:eu-repo/semantics/altIdentifier/doi/10.1007/s12010-011-9277-0info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-17T10:41:40Zoai:ri.conicet.gov.ar:11336/95875instacron: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:34982025-09-17 10:41:40.921CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
title Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
spellingShingle Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
Bruno, Mariela Anahí
BROMELIA HIERONYMI
BROMELIACEAE
CLONING
CYSTEINE ENDOPEPTIDASE
HIERONYMAIN II
PROTEOMIC TOOLS
SEQUENCING
title_short Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
title_full Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
title_fullStr Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
title_full_unstemmed Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
title_sort Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez
dc.creator.none.fl_str_mv Bruno, Mariela Anahí
Trejo, Sebastian Alejandro
Aviles, Francesc Xavier
Caffini, Nestor Oscar
Lopez, Laura Maria Isabel
author Bruno, Mariela Anahí
author_facet Bruno, Mariela Anahí
Trejo, Sebastian Alejandro
Aviles, Francesc Xavier
Caffini, Nestor Oscar
Lopez, Laura Maria Isabel
author_role author
author2 Trejo, Sebastian Alejandro
Aviles, Francesc Xavier
Caffini, Nestor Oscar
Lopez, Laura Maria Isabel
author2_role author
author
author
author
dc.subject.none.fl_str_mv BROMELIA HIERONYMI
BROMELIACEAE
CLONING
CYSTEINE ENDOPEPTIDASE
HIERONYMAIN II
PROTEOMIC TOOLS
SEQUENCING
topic BROMELIA HIERONYMI
BROMELIACEAE
CLONING
CYSTEINE ENDOPEPTIDASE
HIERONYMAIN II
PROTEOMIC TOOLS
SEQUENCING
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Fruits of Bromelia hieronymi, a tropical South American plant, possess a high content of peptidases with potential biotechnological uses. Total RNA was extracted from unripe fruits and peptidase cDNA was obtained by 3′RACE-PCR. The consensus sequence of the cysteine peptidase cDNA contained 875 bp, the 690 first ones codifying for a hypothetical polypeptide chain of the mature peptidase, named Bh-CP1 (molecular mass 24.773 kDa, pI 8.6, extinction molar coefficient 58,705 M−1 cm−1). Bh-CP1 sequence shows a high percentage of identity with those of other cysteine plant proteases. The presence of highly preserved residues is observed, like those forming the catalytic site (Gln19, Cys25, His159, and Asn175, papain numbering), as well as other six Cys residues, involved in the formation of disulfide bounds. Molecular modeling results suggest the enzyme belongs to the α + β class of proteins, with two disulfide bridges (Cys23–Cys63 and Cys57–Cys96) in the α domain, while the β domain is stabilized by another disulfide bridge (Cys153–Cys203). Additionally, peptide mass fingerprints (PMFs) of the three peptidases previously isolated from B. hieronymi fruits (namely hieronymain I, II, and III) were performed and compared with the theoretical fingerprint of PMF of Bh-CP1, showing a partial matching between the in silico-translated protein and hieronymain II.
Fil: Bruno, Mariela Anahí. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Trejo, Sebastian Alejandro. Universidad Autonoma de Barcelona. Departamento de Biología; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Aviles, Francesc Xavier. Universidad Autonoma de Barcelona. Departamento de Biología; España
Fil: Caffini, Nestor Oscar. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina
Fil: Lopez, Laura Maria Isabel. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Fruits of Bromelia hieronymi, a tropical South American plant, possess a high content of peptidases with potential biotechnological uses. Total RNA was extracted from unripe fruits and peptidase cDNA was obtained by 3′RACE-PCR. The consensus sequence of the cysteine peptidase cDNA contained 875 bp, the 690 first ones codifying for a hypothetical polypeptide chain of the mature peptidase, named Bh-CP1 (molecular mass 24.773 kDa, pI 8.6, extinction molar coefficient 58,705 M−1 cm−1). Bh-CP1 sequence shows a high percentage of identity with those of other cysteine plant proteases. The presence of highly preserved residues is observed, like those forming the catalytic site (Gln19, Cys25, His159, and Asn175, papain numbering), as well as other six Cys residues, involved in the formation of disulfide bounds. Molecular modeling results suggest the enzyme belongs to the α + β class of proteins, with two disulfide bridges (Cys23–Cys63 and Cys57–Cys96) in the α domain, while the β domain is stabilized by another disulfide bridge (Cys153–Cys203). Additionally, peptide mass fingerprints (PMFs) of the three peptidases previously isolated from B. hieronymi fruits (namely hieronymain I, II, and III) were performed and compared with the theoretical fingerprint of PMF of Bh-CP1, showing a partial matching between the in silico-translated protein and hieronymain II.
publishDate 2011
dc.date.none.fl_str_mv 2011-09
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/11336/95875
Bruno, Mariela Anahí; Trejo, Sebastian Alejandro; Aviles, Francesc Xavier; Caffini, Nestor Oscar; Lopez, Laura Maria Isabel; Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez; Humana Press; Applied Biochemistry And Biotechnology; 165; 2; 9-2011; 583-593
0273-2289
CONICET Digital
CONICET
url http://hdl.handle.net/11336/95875
identifier_str_mv Bruno, Mariela Anahí; Trejo, Sebastian Alejandro; Aviles, Francesc Xavier; Caffini, Nestor Oscar; Lopez, Laura Maria Isabel; Cloning, sequencing, and identification using proteomic tools of a protease from bromelia hieronymi mez; Humana Press; Applied Biochemistry And Biotechnology; 165; 2; 9-2011; 583-593
0273-2289
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://link.springer.com/article/10.1007/s12010-011-9277-0
info:eu-repo/semantics/altIdentifier/doi/10.1007/s12010-011-9277-0
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
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Humana Press
publisher.none.fl_str_mv Humana Press
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
_version_ 1843605868823707648
score 13.001348

Publicaciones similares