Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner

Autores
Burgardt, Noelia Ines; Schmidt, Andreas; Manns, Annika; Schutkowski, Alexandra; Jahreis, Günther; Lin, Yi Jan; Schulze, Bianca; Masch, Antonia; Lücke, Christian; Weiwad, Matthias
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Recently we have shown that the peptidyl-prolyl cis/trans isomerase parvulin 17 (Par17) interacts with tubulin in a GTPdependent manner, thereby promoting the formation of microtubules. Microtubule assembly is regulated by Ca2+-loaded calmodulin (Ca2+/CaM) both in the intact cell and under in vitro conditions via direct interaction with microtubule-associated proteins. Here we provide the first evidence that Ca2+/CaM interacts also with Par17 in a physiologically relevant way, thus preventing Par17-promoted microtubule assembly. In contrast, parvulin 14 (Par14), which lacks only the first 25 N-terminal residues of the Par17 sequence, does not interact with Ca2+/CaM, indicating that this interaction is exclusive for Par17. Pulldown experiments and chemical shift perturbation analysis with 15N-labeled Par17 furthermore confirmed that calmodulin (CaM) interacts in a Ca2+-dependent manner with the Par17 N terminus. The reverse experiment with 15N-labeled Ca2+/CaM demonstrated that the N-terminal Par17 segment binds to both CaM lobes simultaneously, indicating that Ca2+/CaM undergoes a conformational change to form a binding channel between its two lobes, apparently similar to the structure of the CaM-smMLCK796-815 complex. In vitro tubulin polymerization assays furthermore showed that Ca2+/CaM completely suppresses Par17-promoted microtubule assembly. The results imply that Ca2+/CaM binding to the N-terminal segment of Par17 causes steric hindrance of the Par17 active site, thus interfering with the Par17/tubulin interaction. This Ca2+/CaM-mediated control of Par17-assisted microtubule assembly may provide a mechanism that couples Ca2+ signaling with microtubule function.
Fil: Burgardt, Noelia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Schmidt, Andreas. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Manns, Annika. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Schutkowski, Alexandra. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Jahreis, Günther. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Lin, Yi Jan. Kaohsiung Medical University; Alemania
Fil: Schulze, Bianca. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Masch, Antonia. Martin Luther University Halle Wittenberg; Alemania
Fil: Lücke, Christian. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Weiwad, Matthias. Max Planck Research Unit for Enzymology of Protein Folding; Alemania. Martin Luther University Halle Wittenberg; Alemania
Materia
NUCLEAR MAGNETIC RESONANCE
PROTEIN-PROTEIN INTERACTION
PARVULIN
CALMODULIN
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:11336/67072
Acceder
id CONICETDig_7dd265594e147d0bbe4030f4f996efa0
oai_identifier_str oai:ri.conicet.gov.ar:11336/67072
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent mannerBurgardt, Noelia InesSchmidt, AndreasManns, AnnikaSchutkowski, AlexandraJahreis, GüntherLin, Yi JanSchulze, BiancaMasch, AntoniaLücke, ChristianWeiwad, MatthiasNUCLEAR MAGNETIC RESONANCEPROTEIN-PROTEIN INTERACTIONPARVULINCALMODULINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Recently we have shown that the peptidyl-prolyl cis/trans isomerase parvulin 17 (Par17) interacts with tubulin in a GTPdependent manner, thereby promoting the formation of microtubules. Microtubule assembly is regulated by Ca2+-loaded calmodulin (Ca2+/CaM) both in the intact cell and under in vitro conditions via direct interaction with microtubule-associated proteins. Here we provide the first evidence that Ca2+/CaM interacts also with Par17 in a physiologically relevant way, thus preventing Par17-promoted microtubule assembly. In contrast, parvulin 14 (Par14), which lacks only the first 25 N-terminal residues of the Par17 sequence, does not interact with Ca2+/CaM, indicating that this interaction is exclusive for Par17. Pulldown experiments and chemical shift perturbation analysis with 15N-labeled Par17 furthermore confirmed that calmodulin (CaM) interacts in a Ca2+-dependent manner with the Par17 N terminus. The reverse experiment with 15N-labeled Ca2+/CaM demonstrated that the N-terminal Par17 segment binds to both CaM lobes simultaneously, indicating that Ca2+/CaM undergoes a conformational change to form a binding channel between its two lobes, apparently similar to the structure of the CaM-smMLCK796-815 complex. In vitro tubulin polymerization assays furthermore showed that Ca2+/CaM completely suppresses Par17-promoted microtubule assembly. The results imply that Ca2+/CaM binding to the N-terminal segment of Par17 causes steric hindrance of the Par17 active site, thus interfering with the Par17/tubulin interaction. This Ca2+/CaM-mediated control of Par17-assisted microtubule assembly may provide a mechanism that couples Ca2+ signaling with microtubule function.Fil: Burgardt, Noelia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Max Planck Research Unit for Enzymology of Protein Folding; AlemaniaFil: Schmidt, Andreas. Max Planck Research Unit for Enzymology of Protein Folding; AlemaniaFil: Manns, Annika. Max Planck Research Unit for Enzymology of Protein Folding; AlemaniaFil: Schutkowski, Alexandra. Max Planck Research Unit for Enzymology of Protein Folding; AlemaniaFil: Jahreis, Günther. Max Planck Research Unit for Enzymology of Protein Folding; AlemaniaFil: Lin, Yi Jan. Kaohsiung Medical University; AlemaniaFil: Schulze, Bianca. Max Planck Research Unit for Enzymology of Protein Folding; AlemaniaFil: Masch, Antonia. Martin Luther University Halle Wittenberg; AlemaniaFil: Lücke, Christian. Max Planck Research Unit for Enzymology of Protein Folding; AlemaniaFil: Weiwad, Matthias. Max Planck Research Unit for Enzymology of Protein Folding; Alemania. Martin Luther University Halle Wittenberg; AlemaniaAmerican Society for Biochemistry and Molecular Biology2015-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/67072Burgardt, Noelia Ines; Schmidt, Andreas; Manns, Annika; Schutkowski, Alexandra; Jahreis, Günther; et al.; Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 290; 27; 7-2015; 16708-167220021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/290/27/16708info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M114.593228info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:36:27Zoai:ri.conicet.gov.ar:11336/67072instacron: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:34982025-09-29 09:36:28.002CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
title Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
spellingShingle Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
Burgardt, Noelia Ines
NUCLEAR MAGNETIC RESONANCE
PROTEIN-PROTEIN INTERACTION
PARVULIN
CALMODULIN
title_short Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
title_full Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
title_fullStr Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
title_full_unstemmed Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
title_sort Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner
dc.creator.none.fl_str_mv Burgardt, Noelia Ines
Schmidt, Andreas
Manns, Annika
Schutkowski, Alexandra
Jahreis, Günther
Lin, Yi Jan
Schulze, Bianca
Masch, Antonia
Lücke, Christian
Weiwad, Matthias
author Burgardt, Noelia Ines
author_facet Burgardt, Noelia Ines
Schmidt, Andreas
Manns, Annika
Schutkowski, Alexandra
Jahreis, Günther
Lin, Yi Jan
Schulze, Bianca
Masch, Antonia
Lücke, Christian
Weiwad, Matthias
author_role author
author2 Schmidt, Andreas
Manns, Annika
Schutkowski, Alexandra
Jahreis, Günther
Lin, Yi Jan
Schulze, Bianca
Masch, Antonia
Lücke, Christian
Weiwad, Matthias
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv NUCLEAR MAGNETIC RESONANCE
PROTEIN-PROTEIN INTERACTION
PARVULIN
CALMODULIN
topic NUCLEAR MAGNETIC RESONANCE
PROTEIN-PROTEIN INTERACTION
PARVULIN
CALMODULIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Recently we have shown that the peptidyl-prolyl cis/trans isomerase parvulin 17 (Par17) interacts with tubulin in a GTPdependent manner, thereby promoting the formation of microtubules. Microtubule assembly is regulated by Ca2+-loaded calmodulin (Ca2+/CaM) both in the intact cell and under in vitro conditions via direct interaction with microtubule-associated proteins. Here we provide the first evidence that Ca2+/CaM interacts also with Par17 in a physiologically relevant way, thus preventing Par17-promoted microtubule assembly. In contrast, parvulin 14 (Par14), which lacks only the first 25 N-terminal residues of the Par17 sequence, does not interact with Ca2+/CaM, indicating that this interaction is exclusive for Par17. Pulldown experiments and chemical shift perturbation analysis with 15N-labeled Par17 furthermore confirmed that calmodulin (CaM) interacts in a Ca2+-dependent manner with the Par17 N terminus. The reverse experiment with 15N-labeled Ca2+/CaM demonstrated that the N-terminal Par17 segment binds to both CaM lobes simultaneously, indicating that Ca2+/CaM undergoes a conformational change to form a binding channel between its two lobes, apparently similar to the structure of the CaM-smMLCK796-815 complex. In vitro tubulin polymerization assays furthermore showed that Ca2+/CaM completely suppresses Par17-promoted microtubule assembly. The results imply that Ca2+/CaM binding to the N-terminal segment of Par17 causes steric hindrance of the Par17 active site, thus interfering with the Par17/tubulin interaction. This Ca2+/CaM-mediated control of Par17-assisted microtubule assembly may provide a mechanism that couples Ca2+ signaling with microtubule function.
Fil: Burgardt, Noelia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Schmidt, Andreas. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Manns, Annika. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Schutkowski, Alexandra. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Jahreis, Günther. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Lin, Yi Jan. Kaohsiung Medical University; Alemania
Fil: Schulze, Bianca. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Masch, Antonia. Martin Luther University Halle Wittenberg; Alemania
Fil: Lücke, Christian. Max Planck Research Unit for Enzymology of Protein Folding; Alemania
Fil: Weiwad, Matthias. Max Planck Research Unit for Enzymology of Protein Folding; Alemania. Martin Luther University Halle Wittenberg; Alemania
description Recently we have shown that the peptidyl-prolyl cis/trans isomerase parvulin 17 (Par17) interacts with tubulin in a GTPdependent manner, thereby promoting the formation of microtubules. Microtubule assembly is regulated by Ca2+-loaded calmodulin (Ca2+/CaM) both in the intact cell and under in vitro conditions via direct interaction with microtubule-associated proteins. Here we provide the first evidence that Ca2+/CaM interacts also with Par17 in a physiologically relevant way, thus preventing Par17-promoted microtubule assembly. In contrast, parvulin 14 (Par14), which lacks only the first 25 N-terminal residues of the Par17 sequence, does not interact with Ca2+/CaM, indicating that this interaction is exclusive for Par17. Pulldown experiments and chemical shift perturbation analysis with 15N-labeled Par17 furthermore confirmed that calmodulin (CaM) interacts in a Ca2+-dependent manner with the Par17 N terminus. The reverse experiment with 15N-labeled Ca2+/CaM demonstrated that the N-terminal Par17 segment binds to both CaM lobes simultaneously, indicating that Ca2+/CaM undergoes a conformational change to form a binding channel between its two lobes, apparently similar to the structure of the CaM-smMLCK796-815 complex. In vitro tubulin polymerization assays furthermore showed that Ca2+/CaM completely suppresses Par17-promoted microtubule assembly. The results imply that Ca2+/CaM binding to the N-terminal segment of Par17 causes steric hindrance of the Par17 active site, thus interfering with the Par17/tubulin interaction. This Ca2+/CaM-mediated control of Par17-assisted microtubule assembly may provide a mechanism that couples Ca2+ signaling with microtubule function.
publishDate 2015
dc.date.none.fl_str_mv 2015-07
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/11336/67072
Burgardt, Noelia Ines; Schmidt, Andreas; Manns, Annika; Schutkowski, Alexandra; Jahreis, Günther; et al.; Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 290; 27; 7-2015; 16708-16722
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/67072
identifier_str_mv Burgardt, Noelia Ines; Schmidt, Andreas; Manns, Annika; Schutkowski, Alexandra; Jahreis, Günther; et al.; Parvulin 17-catalyzed tubulin polymerization is regulated by calmodulin in a calcium-dependent manner; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 290; 27; 7-2015; 16708-16722
0021-9258
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.jbc.org/content/290/27/16708
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M114.593228
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 American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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 13.070432

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