Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding

Autores
Camargo, Ana I.; Wiggers, Helton J.; Damalio, Julio C. P.; Araujo, Ana P. U.; Ribichich, Karina Fabiana; Camargo, Paulo C. de
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Centrins are calcium-binding proteins associated with microtubules organizing centers. Members of two divergent subfamilies of centrins were found in the aquatic fungus Blastocladiella emersonii, contrasting with the occurrence of only one member known for the better explored terrestrial fungi. BeCen1, shows greatest identity with human centrins HsCen1, HsCen2 and green algae centrin CrCenp, while BeCen3 records largest identity with human centrin HsCen3 and yeast centrin Cdc31p. Following the discovery of this unique feature, BeCen1 and BeCen3 centrins were produced to study whether these proteins had distinct features upon calcium binding. Circular dichroism showed opposite calcium binding effects on the alpha-helix arrangement of the secondary structure. The spectra indicated a decrease in alpha-helix signal for holo-BeCen1 contrasting with an increase for holo-BeCen3. In addition, only BeCen1 refolds after being denatured. The fluorescence emission of the hydrophobic probe ANS increases for both proteins likely due to hydrophobic exposure, however, only BeCen1 presents a clear blue shift when calcium is added. ITC experiments identified four calcium binding sites for both proteins. In contrast to calcium binding to BeCen1, which is mainly endothermic, binding to BeCen3 is mainly exothermic. Light-scattering evidenced the formation of large particles in solution for BeCen1 and BeCen3 at temperatures above 30°C and 40°C, respectively. Atomic force microscopy confirmed the presence of supramolecular structures, which differ in the compactness and branching degree. Binding of calcium leads to different structural changes in BeCen1 und BeCen3 and the thermodynamic characteristics of the interaction also differ.
Fil: Camargo, Ana I.. Universidade de Sao Paulo; Brasil
Fil: Wiggers, Helton J.. Universidade de Sao Paulo; Brasil
Fil: Damalio, Julio C. P.. Universidade de Sao Paulo; Brasil
Fil: Araujo, Ana P. U.. Universidade de Sao Paulo; Brasil
Fil: Ribichich, Karina Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina. Universidade de Sao Paulo; Brasil
Fil: Camargo, Paulo C. de. Universidade Federal do Paraná; Brasil
Materia
Centrins
Blastocladiella Emersonii
Calcium-Binding Protein
Ef-Hand Protein
Isothermal Titration Calorimetry (Itc)
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/23467
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/23467
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium bindingCamargo, Ana I.Wiggers, Helton J.Damalio, Julio C. P.Araujo, Ana P. U.Ribichich, Karina FabianaCamargo, Paulo C. deCentrinsBlastocladiella EmersoniiCalcium-Binding ProteinEf-Hand ProteinIsothermal Titration Calorimetry (Itc)https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Centrins are calcium-binding proteins associated with microtubules organizing centers. Members of two divergent subfamilies of centrins were found in the aquatic fungus Blastocladiella emersonii, contrasting with the occurrence of only one member known for the better explored terrestrial fungi. BeCen1, shows greatest identity with human centrins HsCen1, HsCen2 and green algae centrin CrCenp, while BeCen3 records largest identity with human centrin HsCen3 and yeast centrin Cdc31p. Following the discovery of this unique feature, BeCen1 and BeCen3 centrins were produced to study whether these proteins had distinct features upon calcium binding. Circular dichroism showed opposite calcium binding effects on the alpha-helix arrangement of the secondary structure. The spectra indicated a decrease in alpha-helix signal for holo-BeCen1 contrasting with an increase for holo-BeCen3. In addition, only BeCen1 refolds after being denatured. The fluorescence emission of the hydrophobic probe ANS increases for both proteins likely due to hydrophobic exposure, however, only BeCen1 presents a clear blue shift when calcium is added. ITC experiments identified four calcium binding sites for both proteins. In contrast to calcium binding to BeCen1, which is mainly endothermic, binding to BeCen3 is mainly exothermic. Light-scattering evidenced the formation of large particles in solution for BeCen1 and BeCen3 at temperatures above 30°C and 40°C, respectively. Atomic force microscopy confirmed the presence of supramolecular structures, which differ in the compactness and branching degree. Binding of calcium leads to different structural changes in BeCen1 und BeCen3 and the thermodynamic characteristics of the interaction also differ.Fil: Camargo, Ana I.. Universidade de Sao Paulo; BrasilFil: Wiggers, Helton J.. Universidade de Sao Paulo; BrasilFil: Damalio, Julio C. P.. Universidade de Sao Paulo; BrasilFil: Araujo, Ana P. U.. Universidade de Sao Paulo; BrasilFil: Ribichich, Karina Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina. Universidade de Sao Paulo; BrasilFil: Camargo, Paulo C. de. Universidade Federal do Paraná; BrasilElsevier2013-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/23467Camargo, Ana I.; Wiggers, Helton J.; Damalio, Julio C. P.; Araujo, Ana P. U.; Ribichich, Karina Fabiana; et al.; Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding; Elsevier; Biochimica Et Biophysica Acta-proteins And Proteomics; 1834; 12; 12-2013; 2823-28311570-9639CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1570963913003658info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbapap.2013.10.007info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:00:56Zoai:ri.conicet.gov.ar:11336/23467instacron: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 10:00:56.469CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
title Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
spellingShingle Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
Camargo, Ana I.
Centrins
Blastocladiella Emersonii
Calcium-Binding Protein
Ef-Hand Protein
Isothermal Titration Calorimetry (Itc)
title_short Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
title_full Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
title_fullStr Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
title_full_unstemmed Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
title_sort Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding
dc.creator.none.fl_str_mv Camargo, Ana I.
Wiggers, Helton J.
Damalio, Julio C. P.
Araujo, Ana P. U.
Ribichich, Karina Fabiana
Camargo, Paulo C. de
author Camargo, Ana I.
author_facet Camargo, Ana I.
Wiggers, Helton J.
Damalio, Julio C. P.
Araujo, Ana P. U.
Ribichich, Karina Fabiana
Camargo, Paulo C. de
author_role author
author2 Wiggers, Helton J.
Damalio, Julio C. P.
Araujo, Ana P. U.
Ribichich, Karina Fabiana
Camargo, Paulo C. de
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Centrins
Blastocladiella Emersonii
Calcium-Binding Protein
Ef-Hand Protein
Isothermal Titration Calorimetry (Itc)
topic Centrins
Blastocladiella Emersonii
Calcium-Binding Protein
Ef-Hand Protein
Isothermal Titration Calorimetry (Itc)
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Centrins are calcium-binding proteins associated with microtubules organizing centers. Members of two divergent subfamilies of centrins were found in the aquatic fungus Blastocladiella emersonii, contrasting with the occurrence of only one member known for the better explored terrestrial fungi. BeCen1, shows greatest identity with human centrins HsCen1, HsCen2 and green algae centrin CrCenp, while BeCen3 records largest identity with human centrin HsCen3 and yeast centrin Cdc31p. Following the discovery of this unique feature, BeCen1 and BeCen3 centrins were produced to study whether these proteins had distinct features upon calcium binding. Circular dichroism showed opposite calcium binding effects on the alpha-helix arrangement of the secondary structure. The spectra indicated a decrease in alpha-helix signal for holo-BeCen1 contrasting with an increase for holo-BeCen3. In addition, only BeCen1 refolds after being denatured. The fluorescence emission of the hydrophobic probe ANS increases for both proteins likely due to hydrophobic exposure, however, only BeCen1 presents a clear blue shift when calcium is added. ITC experiments identified four calcium binding sites for both proteins. In contrast to calcium binding to BeCen1, which is mainly endothermic, binding to BeCen3 is mainly exothermic. Light-scattering evidenced the formation of large particles in solution for BeCen1 and BeCen3 at temperatures above 30°C and 40°C, respectively. Atomic force microscopy confirmed the presence of supramolecular structures, which differ in the compactness and branching degree. Binding of calcium leads to different structural changes in BeCen1 und BeCen3 and the thermodynamic characteristics of the interaction also differ.
Fil: Camargo, Ana I.. Universidade de Sao Paulo; Brasil
Fil: Wiggers, Helton J.. Universidade de Sao Paulo; Brasil
Fil: Damalio, Julio C. P.. Universidade de Sao Paulo; Brasil
Fil: Araujo, Ana P. U.. Universidade de Sao Paulo; Brasil
Fil: Ribichich, Karina Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina. Universidade de Sao Paulo; Brasil
Fil: Camargo, Paulo C. de. Universidade Federal do Paraná; Brasil
description Centrins are calcium-binding proteins associated with microtubules organizing centers. Members of two divergent subfamilies of centrins were found in the aquatic fungus Blastocladiella emersonii, contrasting with the occurrence of only one member known for the better explored terrestrial fungi. BeCen1, shows greatest identity with human centrins HsCen1, HsCen2 and green algae centrin CrCenp, while BeCen3 records largest identity with human centrin HsCen3 and yeast centrin Cdc31p. Following the discovery of this unique feature, BeCen1 and BeCen3 centrins were produced to study whether these proteins had distinct features upon calcium binding. Circular dichroism showed opposite calcium binding effects on the alpha-helix arrangement of the secondary structure. The spectra indicated a decrease in alpha-helix signal for holo-BeCen1 contrasting with an increase for holo-BeCen3. In addition, only BeCen1 refolds after being denatured. The fluorescence emission of the hydrophobic probe ANS increases for both proteins likely due to hydrophobic exposure, however, only BeCen1 presents a clear blue shift when calcium is added. ITC experiments identified four calcium binding sites for both proteins. In contrast to calcium binding to BeCen1, which is mainly endothermic, binding to BeCen3 is mainly exothermic. Light-scattering evidenced the formation of large particles in solution for BeCen1 and BeCen3 at temperatures above 30°C and 40°C, respectively. Atomic force microscopy confirmed the presence of supramolecular structures, which differ in the compactness and branching degree. Binding of calcium leads to different structural changes in BeCen1 und BeCen3 and the thermodynamic characteristics of the interaction also differ.
publishDate 2013
dc.date.none.fl_str_mv 2013-12
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/11336/23467
Camargo, Ana I.; Wiggers, Helton J.; Damalio, Julio C. P.; Araujo, Ana P. U.; Ribichich, Karina Fabiana; et al.; Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding; Elsevier; Biochimica Et Biophysica Acta-proteins And Proteomics; 1834; 12; 12-2013; 2823-2831
1570-9639
CONICET Digital
CONICET
url http://hdl.handle.net/11336/23467
identifier_str_mv Camargo, Ana I.; Wiggers, Helton J.; Damalio, Julio C. P.; Araujo, Ana P. U.; Ribichich, Karina Fabiana; et al.; Structural and thermodynamic studies of two centrin isoforms from Blastocladiella emersonii upon calcium binding; Elsevier; Biochimica Et Biophysica Acta-proteins And Proteomics; 1834; 12; 12-2013; 2823-2831
1570-9639
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1570963913003658
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbapap.2013.10.007
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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 13.13397

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