Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation

Autores
Chaudhuri, Pinaki; Rosenbaum, Michael; Sinharoy, Pritam; Damron, Derek S.; Birnbaumer, Lutz; Graham, Linda
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Lipid oxidation products, including lysophosphatidylcholine (lysoPC), activate canonical transient receptor potential 6 (TRPC6) channels leading to inhibition of endothelial cell (EC) migration in vitro and delayed EC healing of arterial injuries in vivo. The precise mechanism through which lysoPC activates TRPC6 channels is not known, but calmodulin (CaM) contributes to the regulation of TRPC channels. Using site-directed mutagenesis, cDNAs were generated in which Tyr99 or Tyr138 of CaM was replaced with Phe, generating mutant CaM, Phe99-CaM, or Phe138-CaM, respectively. In ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM, but not in ECs transfected with pcDNA3.1-myc-His-Phe138-CaM, the lysoPC-induced TRPC6-CaM dissociation and TRPC6 externalization was disrupted. Also, the lysoPC-induced increase in intracellular calcium concentration was inhibited in ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM. Blocking phosphorylation of CaM at Tyr99 also reduced CaM association with the p85 subunit and subsequent activation of phosphatidylinositol 3-kinase (PI3K). This prevented the increase in phosphatidylinositol (3,4,5)-Trisphosphate (PIP3) and the translocation of TRPC6 to the cell membrane and reduced the inhibition of ECmigration by lysoPC. These findings suggest that lysoPC induces CaM phosphorylation at Tyr99 by a Src family kinase and that phosphorylated CaM activates PI3K to produce PIP3, which promotes TRPC6 translocation to the cell membrane.
Fil: Chaudhuri, Pinaki. Cleveland Clinic; Estados Unidos
Fil: Rosenbaum, Michael. Louis Stokes Cleveland Veterans Affairs Medical Center; Estados Unidos
Fil: Sinharoy, Pritam. Kent State University; Estados Unidos
Fil: Damron, Derek S.. Kent State University; Estados Unidos
Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Graham, Linda. Cleveland Clinic; Estados Unidos
Materia
Endothelial
Calmodulin
Pi3 Kinase
Trpc6
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:11336/62879
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/62879
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activationChaudhuri, PinakiRosenbaum, MichaelSinharoy, PritamDamron, Derek S.Birnbaumer, LutzGraham, LindaEndothelialCalmodulinPi3 KinaseTrpc6https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Lipid oxidation products, including lysophosphatidylcholine (lysoPC), activate canonical transient receptor potential 6 (TRPC6) channels leading to inhibition of endothelial cell (EC) migration in vitro and delayed EC healing of arterial injuries in vivo. The precise mechanism through which lysoPC activates TRPC6 channels is not known, but calmodulin (CaM) contributes to the regulation of TRPC channels. Using site-directed mutagenesis, cDNAs were generated in which Tyr99 or Tyr138 of CaM was replaced with Phe, generating mutant CaM, Phe99-CaM, or Phe138-CaM, respectively. In ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM, but not in ECs transfected with pcDNA3.1-myc-His-Phe138-CaM, the lysoPC-induced TRPC6-CaM dissociation and TRPC6 externalization was disrupted. Also, the lysoPC-induced increase in intracellular calcium concentration was inhibited in ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM. Blocking phosphorylation of CaM at Tyr99 also reduced CaM association with the p85 subunit and subsequent activation of phosphatidylinositol 3-kinase (PI3K). This prevented the increase in phosphatidylinositol (3,4,5)-Trisphosphate (PIP3) and the translocation of TRPC6 to the cell membrane and reduced the inhibition of ECmigration by lysoPC. These findings suggest that lysoPC induces CaM phosphorylation at Tyr99 by a Src family kinase and that phosphorylated CaM activates PI3K to produce PIP3, which promotes TRPC6 translocation to the cell membrane.Fil: Chaudhuri, Pinaki. Cleveland Clinic; Estados UnidosFil: Rosenbaum, Michael. Louis Stokes Cleveland Veterans Affairs Medical Center; Estados UnidosFil: Sinharoy, Pritam. Kent State University; Estados UnidosFil: Damron, Derek S.. Kent State University; Estados UnidosFil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Graham, Linda. Cleveland Clinic; Estados UnidosNational Academy of Sciences2016-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/62879Chaudhuri, Pinaki; Rosenbaum, Michael; Sinharoy, Pritam; Damron, Derek S.; Birnbaumer, Lutz; et al.; Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 113; 8; 2-2016; 2110-21150027-8424CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.pnas.org/content/113/8/2110info:eu-repo/semantics/altIdentifier/doi/10.1073/pnas.1600371113info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:47:04Zoai:ri.conicet.gov.ar:11336/62879instacron: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 09:47:04.264CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
title Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
spellingShingle Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
Chaudhuri, Pinaki
Endothelial
Calmodulin
Pi3 Kinase
Trpc6
title_short Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
title_full Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
title_fullStr Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
title_full_unstemmed Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
title_sort Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation
dc.creator.none.fl_str_mv Chaudhuri, Pinaki
Rosenbaum, Michael
Sinharoy, Pritam
Damron, Derek S.
Birnbaumer, Lutz
Graham, Linda
author Chaudhuri, Pinaki
author_facet Chaudhuri, Pinaki
Rosenbaum, Michael
Sinharoy, Pritam
Damron, Derek S.
Birnbaumer, Lutz
Graham, Linda
author_role author
author2 Rosenbaum, Michael
Sinharoy, Pritam
Damron, Derek S.
Birnbaumer, Lutz
Graham, Linda
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Endothelial
Calmodulin
Pi3 Kinase
Trpc6
topic Endothelial
Calmodulin
Pi3 Kinase
Trpc6
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Lipid oxidation products, including lysophosphatidylcholine (lysoPC), activate canonical transient receptor potential 6 (TRPC6) channels leading to inhibition of endothelial cell (EC) migration in vitro and delayed EC healing of arterial injuries in vivo. The precise mechanism through which lysoPC activates TRPC6 channels is not known, but calmodulin (CaM) contributes to the regulation of TRPC channels. Using site-directed mutagenesis, cDNAs were generated in which Tyr99 or Tyr138 of CaM was replaced with Phe, generating mutant CaM, Phe99-CaM, or Phe138-CaM, respectively. In ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM, but not in ECs transfected with pcDNA3.1-myc-His-Phe138-CaM, the lysoPC-induced TRPC6-CaM dissociation and TRPC6 externalization was disrupted. Also, the lysoPC-induced increase in intracellular calcium concentration was inhibited in ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM. Blocking phosphorylation of CaM at Tyr99 also reduced CaM association with the p85 subunit and subsequent activation of phosphatidylinositol 3-kinase (PI3K). This prevented the increase in phosphatidylinositol (3,4,5)-Trisphosphate (PIP3) and the translocation of TRPC6 to the cell membrane and reduced the inhibition of ECmigration by lysoPC. These findings suggest that lysoPC induces CaM phosphorylation at Tyr99 by a Src family kinase and that phosphorylated CaM activates PI3K to produce PIP3, which promotes TRPC6 translocation to the cell membrane.
Fil: Chaudhuri, Pinaki. Cleveland Clinic; Estados Unidos
Fil: Rosenbaum, Michael. Louis Stokes Cleveland Veterans Affairs Medical Center; Estados Unidos
Fil: Sinharoy, Pritam. Kent State University; Estados Unidos
Fil: Damron, Derek S.. Kent State University; Estados Unidos
Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Graham, Linda. Cleveland Clinic; Estados Unidos
description Lipid oxidation products, including lysophosphatidylcholine (lysoPC), activate canonical transient receptor potential 6 (TRPC6) channels leading to inhibition of endothelial cell (EC) migration in vitro and delayed EC healing of arterial injuries in vivo. The precise mechanism through which lysoPC activates TRPC6 channels is not known, but calmodulin (CaM) contributes to the regulation of TRPC channels. Using site-directed mutagenesis, cDNAs were generated in which Tyr99 or Tyr138 of CaM was replaced with Phe, generating mutant CaM, Phe99-CaM, or Phe138-CaM, respectively. In ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM, but not in ECs transfected with pcDNA3.1-myc-His-Phe138-CaM, the lysoPC-induced TRPC6-CaM dissociation and TRPC6 externalization was disrupted. Also, the lysoPC-induced increase in intracellular calcium concentration was inhibited in ECs transiently transfected with pcDNA3.1-myc-His-Phe99-CaM. Blocking phosphorylation of CaM at Tyr99 also reduced CaM association with the p85 subunit and subsequent activation of phosphatidylinositol 3-kinase (PI3K). This prevented the increase in phosphatidylinositol (3,4,5)-Trisphosphate (PIP3) and the translocation of TRPC6 to the cell membrane and reduced the inhibition of ECmigration by lysoPC. These findings suggest that lysoPC induces CaM phosphorylation at Tyr99 by a Src family kinase and that phosphorylated CaM activates PI3K to produce PIP3, which promotes TRPC6 translocation to the cell membrane.
publishDate 2016
dc.date.none.fl_str_mv 2016-02
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/11336/62879
Chaudhuri, Pinaki; Rosenbaum, Michael; Sinharoy, Pritam; Damron, Derek S.; Birnbaumer, Lutz; et al.; Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 113; 8; 2-2016; 2110-2115
0027-8424
CONICET Digital
CONICET
url http://hdl.handle.net/11336/62879
identifier_str_mv Chaudhuri, Pinaki; Rosenbaum, Michael; Sinharoy, Pritam; Damron, Derek S.; Birnbaumer, Lutz; et al.; Membrane translocation of TRPC6 channels and endothelial migration are regulated by calmodulin and PI3 kinase activation; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 113; 8; 2-2016; 2110-2115
0027-8424
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.pnas.org/content/113/8/2110
info:eu-repo/semantics/altIdentifier/doi/10.1073/pnas.1600371113
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
dc.publisher.none.fl_str_mv National Academy of Sciences
publisher.none.fl_str_mv National Academy of Sciences
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_ 1842268834932719616
score 13.13397

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