Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family

Autores
Sciorra, Marcelo Daniel; Fantino, Elisa Inés; Giammaria, Veronica; Ulloa, Rita Maria
Año de publicación
2017
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
As sessile organisms, plants have developed complex signal transduction pathways to cope with the environmental fluctuations to which they are exposed during their life cycle. Calcium is a ubiquitous messenger involved in the signaling of environmental stimuli, whose oscillations are decoded by different sensors. Calcium-dependent protein kinases (CDPKs) are key components of calcium regulated signaling cascades in plants. In this work, isoforms StCDPK22 and StCDPK24 from the CDPK family of Solanum tuberosum were characterized. Both isoforms belong to Group III, and contain only three EF-hand sites in their calmodulin-like regulatory domain (CLD) instead of four sites like all other members of the family. StCDPK22 encodes a 59.4 kDa protein (524 aa; pI 7.6) while StCDPK24 encodes a 60 kDa protein (532 aa; pI 6.12), both with an N-terminal variable domain (NTV) which presents myristoylation and palmitoylation consensus sites. StCDPK22 gene (circa 4 kb) is localized in chromosome 10, while StCDPK24 gene (circa 5 kb) is localized in chromosome 11. Both isoforms share the eight exons and seven introns structure and have 80% identity and 89% similarity at the protein level. According to RNAseq data from the potato genome StCDPK22 and StCDPK24 expression is ubiquitous, however StCDPK22 transcripts are much more abundant in stolons, leaves, shoots, tubers and roots. The recombinant 6xHis:StCDPK22 and 6xHis:StCDPK24 were obtained and kinase assays were performed. Although both isoforms are active kinases, StCDPK24 activity is ten-fold higher. Kinetic parameters were determined for StCDPK24. We observed that this kinase can use either Mg2+ or Mn2+ as cofactor and full activity was obtained with 1µM calcium, however kinase activity was also observed in the presence of 10 mM EGTA suggesting that calcium may not be a prerequisite for activity. However, in vitro assays indicate that calcium is required for autophosphorylation.
Fil: Sciorra, Marcelo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Fantino, Elisa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Giammaria, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Ulloa, Rita Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Reunión Conjunta de Sociedades de Biociencias; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Sociedad Argentina de Inmunología
Sociedad Argentina de Andrología
Sociedad Argentina de Biofísica
Sociedad Argentina de Biología
Sociedad Argentina de Farmacología Experimental
Sociedad Argentina de Fisiología
Sociedad Argentina de Hematología
Sociedad Argentina de Protozoología
Materia
CDPK
KINASE ACTIVITY
POTATO
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:11336/235916
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/235916
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) familySciorra, Marcelo DanielFantino, Elisa InésGiammaria, VeronicaUlloa, Rita MariaCDPKKINASE ACTIVITYPOTATOhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1As sessile organisms, plants have developed complex signal transduction pathways to cope with the environmental fluctuations to which they are exposed during their life cycle. Calcium is a ubiquitous messenger involved in the signaling of environmental stimuli, whose oscillations are decoded by different sensors. Calcium-dependent protein kinases (CDPKs) are key components of calcium regulated signaling cascades in plants. In this work, isoforms StCDPK22 and StCDPK24 from the CDPK family of Solanum tuberosum were characterized. Both isoforms belong to Group III, and contain only three EF-hand sites in their calmodulin-like regulatory domain (CLD) instead of four sites like all other members of the family. StCDPK22 encodes a 59.4 kDa protein (524 aa; pI 7.6) while StCDPK24 encodes a 60 kDa protein (532 aa; pI 6.12), both with an N-terminal variable domain (NTV) which presents myristoylation and palmitoylation consensus sites. StCDPK22 gene (circa 4 kb) is localized in chromosome 10, while StCDPK24 gene (circa 5 kb) is localized in chromosome 11. Both isoforms share the eight exons and seven introns structure and have 80% identity and 89% similarity at the protein level. According to RNAseq data from the potato genome StCDPK22 and StCDPK24 expression is ubiquitous, however StCDPK22 transcripts are much more abundant in stolons, leaves, shoots, tubers and roots. The recombinant 6xHis:StCDPK22 and 6xHis:StCDPK24 were obtained and kinase assays were performed. Although both isoforms are active kinases, StCDPK24 activity is ten-fold higher. Kinetic parameters were determined for StCDPK24. We observed that this kinase can use either Mg2+ or Mn2+ as cofactor and full activity was obtained with 1µM calcium, however kinase activity was also observed in the presence of 10 mM EGTA suggesting that calcium may not be a prerequisite for activity. However, in vitro assays indicate that calcium is required for autophosphorylation.Fil: Sciorra, Marcelo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Fantino, Elisa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Giammaria, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Ulloa, Rita Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaReunión Conjunta de Sociedades de Biociencias; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de ProtozoologíaCiudad Autónoma de Buenos AiresArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de Investigación Bioquímica y Biología MolecularSociedad Argentina de InmunologíaSociedad Argentina de AndrologíaSociedad Argentina de BiofísicaSociedad Argentina de BiologíaSociedad Argentina de Farmacología ExperimentalSociedad Argentina de FisiologíaSociedad Argentina de HematologíaSociedad Argentina de ProtozoologíaFundación Revista Medicina2017info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/235916Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family; Reunión Conjunta de Sociedades de Biociencias; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Ciudad Autónoma de Buenos Aires; Argentina; 2017; 301-3011669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://medicinabuenosaires.com/revistas/vol77-17/Vol.77SuplementoI-2017.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:07:50Zoai:ri.conicet.gov.ar:11336/235916instacron: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:34982025-09-03 10:07:51.042CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
title Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
spellingShingle Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
Sciorra, Marcelo Daniel
CDPK
KINASE ACTIVITY
POTATO
title_short Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
title_full Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
title_fullStr Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
title_full_unstemmed Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
title_sort Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family
dc.creator.none.fl_str_mv Sciorra, Marcelo Daniel
Fantino, Elisa Inés
Giammaria, Veronica
Ulloa, Rita Maria
author Sciorra, Marcelo Daniel
author_facet Sciorra, Marcelo Daniel
Fantino, Elisa Inés
Giammaria, Veronica
Ulloa, Rita Maria
author_role author
author2 Fantino, Elisa Inés
Giammaria, Veronica
Ulloa, Rita Maria
author2_role author
author
author
dc.subject.none.fl_str_mv CDPK
KINASE ACTIVITY
POTATO
topic CDPK
KINASE ACTIVITY
POTATO
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv As sessile organisms, plants have developed complex signal transduction pathways to cope with the environmental fluctuations to which they are exposed during their life cycle. Calcium is a ubiquitous messenger involved in the signaling of environmental stimuli, whose oscillations are decoded by different sensors. Calcium-dependent protein kinases (CDPKs) are key components of calcium regulated signaling cascades in plants. In this work, isoforms StCDPK22 and StCDPK24 from the CDPK family of Solanum tuberosum were characterized. Both isoforms belong to Group III, and contain only three EF-hand sites in their calmodulin-like regulatory domain (CLD) instead of four sites like all other members of the family. StCDPK22 encodes a 59.4 kDa protein (524 aa; pI 7.6) while StCDPK24 encodes a 60 kDa protein (532 aa; pI 6.12), both with an N-terminal variable domain (NTV) which presents myristoylation and palmitoylation consensus sites. StCDPK22 gene (circa 4 kb) is localized in chromosome 10, while StCDPK24 gene (circa 5 kb) is localized in chromosome 11. Both isoforms share the eight exons and seven introns structure and have 80% identity and 89% similarity at the protein level. According to RNAseq data from the potato genome StCDPK22 and StCDPK24 expression is ubiquitous, however StCDPK22 transcripts are much more abundant in stolons, leaves, shoots, tubers and roots. The recombinant 6xHis:StCDPK22 and 6xHis:StCDPK24 were obtained and kinase assays were performed. Although both isoforms are active kinases, StCDPK24 activity is ten-fold higher. Kinetic parameters were determined for StCDPK24. We observed that this kinase can use either Mg2+ or Mn2+ as cofactor and full activity was obtained with 1µM calcium, however kinase activity was also observed in the presence of 10 mM EGTA suggesting that calcium may not be a prerequisite for activity. However, in vitro assays indicate that calcium is required for autophosphorylation.
Fil: Sciorra, Marcelo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Fantino, Elisa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Giammaria, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Ulloa, Rita Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Reunión Conjunta de Sociedades de Biociencias; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Sociedad Argentina de Inmunología
Sociedad Argentina de Andrología
Sociedad Argentina de Biofísica
Sociedad Argentina de Biología
Sociedad Argentina de Farmacología Experimental
Sociedad Argentina de Fisiología
Sociedad Argentina de Hematología
Sociedad Argentina de Protozoología
description As sessile organisms, plants have developed complex signal transduction pathways to cope with the environmental fluctuations to which they are exposed during their life cycle. Calcium is a ubiquitous messenger involved in the signaling of environmental stimuli, whose oscillations are decoded by different sensors. Calcium-dependent protein kinases (CDPKs) are key components of calcium regulated signaling cascades in plants. In this work, isoforms StCDPK22 and StCDPK24 from the CDPK family of Solanum tuberosum were characterized. Both isoforms belong to Group III, and contain only three EF-hand sites in their calmodulin-like regulatory domain (CLD) instead of four sites like all other members of the family. StCDPK22 encodes a 59.4 kDa protein (524 aa; pI 7.6) while StCDPK24 encodes a 60 kDa protein (532 aa; pI 6.12), both with an N-terminal variable domain (NTV) which presents myristoylation and palmitoylation consensus sites. StCDPK22 gene (circa 4 kb) is localized in chromosome 10, while StCDPK24 gene (circa 5 kb) is localized in chromosome 11. Both isoforms share the eight exons and seven introns structure and have 80% identity and 89% similarity at the protein level. According to RNAseq data from the potato genome StCDPK22 and StCDPK24 expression is ubiquitous, however StCDPK22 transcripts are much more abundant in stolons, leaves, shoots, tubers and roots. The recombinant 6xHis:StCDPK22 and 6xHis:StCDPK24 were obtained and kinase assays were performed. Although both isoforms are active kinases, StCDPK24 activity is ten-fold higher. Kinetic parameters were determined for StCDPK24. We observed that this kinase can use either Mg2+ or Mn2+ as cofactor and full activity was obtained with 1µM calcium, however kinase activity was also observed in the presence of 10 mM EGTA suggesting that calcium may not be a prerequisite for activity. However, in vitro assays indicate that calcium is required for autophosphorylation.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/235916
Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family; Reunión Conjunta de Sociedades de Biociencias; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Ciudad Autónoma de Buenos Aires; Argentina; 2017; 301-301
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/235916
identifier_str_mv Biochemical characterization of two group III-members of the Solanum tuberosum calcium dependent protein kinase (CDPK) family; Reunión Conjunta de Sociedades de Biociencias; LXII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica; XIX Reunión Anual de la Sociedad Argentina de Biología; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Ciudad Autónoma de Buenos Aires; Argentina; 2017; 301-301
1669-9106
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://medicinabuenosaires.com/revistas/vol77-17/Vol.77SuplementoI-2017.pdf
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https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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reponame_str CONICET Digital (CONICET)
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instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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