Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. C...

Autores
Cotabarren, Juliana; Tellechea, Mariana Edith; Tanco, Sebastián Martín; Lorenzo, Julia; García Pardo, Javier; Avilés, Francesc Xavier; Obregón, Walter David
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Cystine-knot miniproteins (CKMPs) are an intriguing group of cysteine-rich molecules that combine the characteristics of proteins and peptides. Typically, CKMPs are fewer than 50 residues in length and share a characteristic knotted scaffold characterized by the presence of three intramolecular disulfide bonds that form the singular knotted structure. The knot scaffold confers on these proteins remarkable chemical, thermal, and proteolytic stability. Recently, CKMPs have emerged as a novel class of natural molecules with interesting pharmacological properties. In the present work, a novel cystine-knot metallocarboxypeptidase inhibitor (chuPCI) was isolated from tubers of Solanum tuberosum, subsp. andigenum cv. Churqueña. Our results demonstrated that chuPCI is a member of the A/B-type family of metallocarboxypeptidases inhibitors. chuPCI was expressed and characterized by a combination of biochemical and mass spectrometric techniques. Direct comparison of the MALDI-TOF mass spectra for the native and recombinant molecules allowed us to confirm the presence of four different forms of chuPCI in the tubers. The majority of such forms have a molecular weight of 4309 Da and contain a cyclized Gln in the N-terminus. The other three forms are derived from N-terminal and/or C-terminal proteolytic cleavages. Taken together, our results contribute to increase the current repertoire of natural CKMPs.
Facultad de Ciencias Exactas
Materia
Biología
cystine-knot miniproteins; carboxypeptidase inhibitor; plant inhibitor; Solanum tuberosum; protease; Andean potatoes
Cistina
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:10915/68045

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spelling Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. ChurqueñaCotabarren, JulianaTellechea, Mariana EdithTanco, Sebastián MartínLorenzo, JuliaGarcía Pardo, JavierAvilés, Francesc XavierObregón, Walter DavidBiologíacystine-knot miniproteins; carboxypeptidase inhibitor; plant inhibitor; Solanum tuberosum; protease; Andean potatoesCistinaCystine-knot miniproteins (CKMPs) are an intriguing group of cysteine-rich molecules that combine the characteristics of proteins and peptides. Typically, CKMPs are fewer than 50 residues in length and share a characteristic knotted scaffold characterized by the presence of three intramolecular disulfide bonds that form the singular knotted structure. The knot scaffold confers on these proteins remarkable chemical, thermal, and proteolytic stability. Recently, CKMPs have emerged as a novel class of natural molecules with interesting pharmacological properties. In the present work, a novel cystine-knot metallocarboxypeptidase inhibitor (chuPCI) was isolated from tubers of Solanum tuberosum, subsp. andigenum cv. Churqueña. Our results demonstrated that chuPCI is a member of the A/B-type family of metallocarboxypeptidases inhibitors. chuPCI was expressed and characterized by a combination of biochemical and mass spectrometric techniques. Direct comparison of the MALDI-TOF mass spectra for the native and recombinant molecules allowed us to confirm the presence of four different forms of chuPCI in the tubers. The majority of such forms have a molecular weight of 4309 Da and contain a cyclized Gln in the N-terminus. The other three forms are derived from N-terminal and/or C-terminal proteolytic cleavages. Taken together, our results contribute to increase the current repertoire of natural CKMPs.Facultad de Ciencias Exactas2018info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://sedici.unlp.edu.ar/handle/10915/68045enginfo:eu-repo/semantics/altIdentifier/issn/1661-6596info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms19030678info: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:UNLP2025-09-29T11:10:30Zoai:sedici.unlp.edu.ar:10915/68045Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:10:30.73SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
title Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
spellingShingle Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
Cotabarren, Juliana
Biología
cystine-knot miniproteins; carboxypeptidase inhibitor; plant inhibitor; Solanum tuberosum; protease; Andean potatoes
Cistina
title_short Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
title_full Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
title_fullStr Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
title_full_unstemmed Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
title_sort Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from <i>Solanum tuberosum</i> subsp. <i>andigenum</i> cv. Churqueña
dc.creator.none.fl_str_mv Cotabarren, Juliana
Tellechea, Mariana Edith
Tanco, Sebastián Martín
Lorenzo, Julia
García Pardo, Javier
Avilés, Francesc Xavier
Obregón, Walter David
author Cotabarren, Juliana
author_facet Cotabarren, Juliana
Tellechea, Mariana Edith
Tanco, Sebastián Martín
Lorenzo, Julia
García Pardo, Javier
Avilés, Francesc Xavier
Obregón, Walter David
author_role author
author2 Tellechea, Mariana Edith
Tanco, Sebastián Martín
Lorenzo, Julia
García Pardo, Javier
Avilés, Francesc Xavier
Obregón, Walter David
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Biología
cystine-knot miniproteins; carboxypeptidase inhibitor; plant inhibitor; Solanum tuberosum; protease; Andean potatoes
Cistina
topic Biología
cystine-knot miniproteins; carboxypeptidase inhibitor; plant inhibitor; Solanum tuberosum; protease; Andean potatoes
Cistina
dc.description.none.fl_txt_mv Cystine-knot miniproteins (CKMPs) are an intriguing group of cysteine-rich molecules that combine the characteristics of proteins and peptides. Typically, CKMPs are fewer than 50 residues in length and share a characteristic knotted scaffold characterized by the presence of three intramolecular disulfide bonds that form the singular knotted structure. The knot scaffold confers on these proteins remarkable chemical, thermal, and proteolytic stability. Recently, CKMPs have emerged as a novel class of natural molecules with interesting pharmacological properties. In the present work, a novel cystine-knot metallocarboxypeptidase inhibitor (chuPCI) was isolated from tubers of Solanum tuberosum, subsp. andigenum cv. Churqueña. Our results demonstrated that chuPCI is a member of the A/B-type family of metallocarboxypeptidases inhibitors. chuPCI was expressed and characterized by a combination of biochemical and mass spectrometric techniques. Direct comparison of the MALDI-TOF mass spectra for the native and recombinant molecules allowed us to confirm the presence of four different forms of chuPCI in the tubers. The majority of such forms have a molecular weight of 4309 Da and contain a cyclized Gln in the N-terminus. The other three forms are derived from N-terminal and/or C-terminal proteolytic cleavages. Taken together, our results contribute to increase the current repertoire of natural CKMPs.
Facultad de Ciencias Exactas
description Cystine-knot miniproteins (CKMPs) are an intriguing group of cysteine-rich molecules that combine the characteristics of proteins and peptides. Typically, CKMPs are fewer than 50 residues in length and share a characteristic knotted scaffold characterized by the presence of three intramolecular disulfide bonds that form the singular knotted structure. The knot scaffold confers on these proteins remarkable chemical, thermal, and proteolytic stability. Recently, CKMPs have emerged as a novel class of natural molecules with interesting pharmacological properties. In the present work, a novel cystine-knot metallocarboxypeptidase inhibitor (chuPCI) was isolated from tubers of Solanum tuberosum, subsp. andigenum cv. Churqueña. Our results demonstrated that chuPCI is a member of the A/B-type family of metallocarboxypeptidases inhibitors. chuPCI was expressed and characterized by a combination of biochemical and mass spectrometric techniques. Direct comparison of the MALDI-TOF mass spectra for the native and recombinant molecules allowed us to confirm the presence of four different forms of chuPCI in the tubers. The majority of such forms have a molecular weight of 4309 Da and contain a cyclized Gln in the N-terminus. The other three forms are derived from N-terminal and/or C-terminal proteolytic cleavages. Taken together, our results contribute to increase the current repertoire of natural CKMPs.
publishDate 2018
dc.date.none.fl_str_mv 2018
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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status_str publishedVersion
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language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/1661-6596
info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms19030678
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)
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