A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)

Autores
Gross, Lissy Zoe Florens; Sacerdoti, Mariana; Leroux, Alejandro Ezequiel; Rinaldi, Jimena Julieta; Capellari, M. V.; Aramendia, Pedro Francisco; Ghode, A.; Anand, G. S.; Klinke, Sebastian; Biondi, Ricardo Miguel
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Phosphoinositide-dependent protein kinase 1 (PDK1) is a master AGC kinase that phosphorylates at least other 23 AGC kinases, being PKB/Aktthe most relevant substrate downstream of PI3-kinase, important for growth and cell survival and a drug target for cancer treatment. Over the years, our laboratory studied and characterized in detail the catalytic domain of PDK1, as well as the selective activation of substrates such as SGK or S6K, which in order to be phosphorylated require a docking interaction with a hydrophobic site in PDK1 termed the PIF-Pocket. However, this is not the case of Akt/PKB, since it can be activated in a PIF-Pocket independent way. On the other hand, up to date little is known about the mechanistic and structural details of PDK1 full length. Therefore, we are using an interdisciplinary approach to understand how the full length protein is regulated and how this regulation mechanism can be manipulated to specifically inhibit the activation of PKB/Akt. As a result of a medium-scale screening of small compounds we carried out using AlphaScreen technology, we validated a series of small compounds "hits" that modulate PDK1 structure by interaction at different sites on PDK1. We performed hydrogen/deuterium exchange (HDX) experimentsand crystallization screenings to understand the structure of full length PDK1 and how it can be modulated with small compounds. We here present a series of results obtained using HDX on full length PDK1 and the crystal structure of the catalytic domain of PDK1 bound to a small compound identified in our screening. We also present our in vitro studies to understand the oligomerization of PDK1 visualizing single particles using STORM fluorescence microscopy. Finally, we integrate our data to present an updated model on the molecular mechanism of regulation of PDK1.
Fil: Gross, Lissy Zoe Florens. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Sacerdoti, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Leroux, Alejandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Rinaldi, Jimena Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Capellari, M. V.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; Argentina
Fil: Aramendia, Pedro Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; Argentina
Fil: Ghode, A.. National University; Singapur
Fil: Anand, G. S.. National University; Singapur
Fil: Klinke, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Biondi, Ricardo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
LV Reunión Científica anual SAIB y XIV congreso de PABMB
Salta
Argentina
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Pan-American Association For Biochemistry And Molecular Biology
Materia
Protein Kinase
PDK1
Structural Biology
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:11336/155081
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/155081
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)Gross, Lissy Zoe FlorensSacerdoti, MarianaLeroux, Alejandro EzequielRinaldi, Jimena JulietaCapellari, M. V.Aramendia, Pedro FranciscoGhode, A.Anand, G. S.Klinke, SebastianBiondi, Ricardo MiguelProtein KinasePDK1Structural Biologyhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Phosphoinositide-dependent protein kinase 1 (PDK1) is a master AGC kinase that phosphorylates at least other 23 AGC kinases, being PKB/Aktthe most relevant substrate downstream of PI3-kinase, important for growth and cell survival and a drug target for cancer treatment. Over the years, our laboratory studied and characterized in detail the catalytic domain of PDK1, as well as the selective activation of substrates such as SGK or S6K, which in order to be phosphorylated require a docking interaction with a hydrophobic site in PDK1 termed the PIF-Pocket. However, this is not the case of Akt/PKB, since it can be activated in a PIF-Pocket independent way. On the other hand, up to date little is known about the mechanistic and structural details of PDK1 full length. Therefore, we are using an interdisciplinary approach to understand how the full length protein is regulated and how this regulation mechanism can be manipulated to specifically inhibit the activation of PKB/Akt. As a result of a medium-scale screening of small compounds we carried out using AlphaScreen technology, we validated a series of small compounds "hits" that modulate PDK1 structure by interaction at different sites on PDK1. We performed hydrogen/deuterium exchange (HDX) experimentsand crystallization screenings to understand the structure of full length PDK1 and how it can be modulated with small compounds. We here present a series of results obtained using HDX on full length PDK1 and the crystal structure of the catalytic domain of PDK1 bound to a small compound identified in our screening. We also present our in vitro studies to understand the oligomerization of PDK1 visualizing single particles using STORM fluorescence microscopy. Finally, we integrate our data to present an updated model on the molecular mechanism of regulation of PDK1.Fil: Gross, Lissy Zoe Florens. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Sacerdoti, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Leroux, Alejandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Rinaldi, Jimena Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Capellari, M. V.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; ArgentinaFil: Aramendia, Pedro Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; ArgentinaFil: Ghode, A.. National University; SingapurFil: Anand, G. S.. National University; SingapurFil: Klinke, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Biondi, Ricardo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaLV Reunión Científica anual SAIB y XIV congreso de PABMBSaltaArgentinaSociedad Argentina de Investigación Bioquímica y Biología MolecularPan-American Association For Biochemistry And Molecular BiologyBiocell2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/155081A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1); LV Reunión Científica anual SAIB y XIV congreso de PABMB; Salta; Argentina; 2019; 95-950327-9545CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://tesorerasaib.wixsite.com/pabmb-saib2019Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:09:11Zoai:ri.conicet.gov.ar:11336/155081instacron: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 10:09:11.745CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
title A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
spellingShingle A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
Gross, Lissy Zoe Florens
Protein Kinase
PDK1
Structural Biology
title_short A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
title_full A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
title_fullStr A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
title_full_unstemmed A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
title_sort A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1)
dc.creator.none.fl_str_mv Gross, Lissy Zoe Florens
Sacerdoti, Mariana
Leroux, Alejandro Ezequiel
Rinaldi, Jimena Julieta
Capellari, M. V.
Aramendia, Pedro Francisco
Ghode, A.
Anand, G. S.
Klinke, Sebastian
Biondi, Ricardo Miguel
author Gross, Lissy Zoe Florens
author_facet Gross, Lissy Zoe Florens
Sacerdoti, Mariana
Leroux, Alejandro Ezequiel
Rinaldi, Jimena Julieta
Capellari, M. V.
Aramendia, Pedro Francisco
Ghode, A.
Anand, G. S.
Klinke, Sebastian
Biondi, Ricardo Miguel
author_role author
author2 Sacerdoti, Mariana
Leroux, Alejandro Ezequiel
Rinaldi, Jimena Julieta
Capellari, M. V.
Aramendia, Pedro Francisco
Ghode, A.
Anand, G. S.
Klinke, Sebastian
Biondi, Ricardo Miguel
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Protein Kinase
PDK1
Structural Biology
topic Protein Kinase
PDK1
Structural Biology
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Phosphoinositide-dependent protein kinase 1 (PDK1) is a master AGC kinase that phosphorylates at least other 23 AGC kinases, being PKB/Aktthe most relevant substrate downstream of PI3-kinase, important for growth and cell survival and a drug target for cancer treatment. Over the years, our laboratory studied and characterized in detail the catalytic domain of PDK1, as well as the selective activation of substrates such as SGK or S6K, which in order to be phosphorylated require a docking interaction with a hydrophobic site in PDK1 termed the PIF-Pocket. However, this is not the case of Akt/PKB, since it can be activated in a PIF-Pocket independent way. On the other hand, up to date little is known about the mechanistic and structural details of PDK1 full length. Therefore, we are using an interdisciplinary approach to understand how the full length protein is regulated and how this regulation mechanism can be manipulated to specifically inhibit the activation of PKB/Akt. As a result of a medium-scale screening of small compounds we carried out using AlphaScreen technology, we validated a series of small compounds "hits" that modulate PDK1 structure by interaction at different sites on PDK1. We performed hydrogen/deuterium exchange (HDX) experimentsand crystallization screenings to understand the structure of full length PDK1 and how it can be modulated with small compounds. We here present a series of results obtained using HDX on full length PDK1 and the crystal structure of the catalytic domain of PDK1 bound to a small compound identified in our screening. We also present our in vitro studies to understand the oligomerization of PDK1 visualizing single particles using STORM fluorescence microscopy. Finally, we integrate our data to present an updated model on the molecular mechanism of regulation of PDK1.
Fil: Gross, Lissy Zoe Florens. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Sacerdoti, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Leroux, Alejandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Rinaldi, Jimena Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Capellari, M. V.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; Argentina
Fil: Aramendia, Pedro Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; Argentina
Fil: Ghode, A.. National University; Singapur
Fil: Anand, G. S.. National University; Singapur
Fil: Klinke, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Biondi, Ricardo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
LV Reunión Científica anual SAIB y XIV congreso de PABMB
Salta
Argentina
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Pan-American Association For Biochemistry And Molecular Biology
description Phosphoinositide-dependent protein kinase 1 (PDK1) is a master AGC kinase that phosphorylates at least other 23 AGC kinases, being PKB/Aktthe most relevant substrate downstream of PI3-kinase, important for growth and cell survival and a drug target for cancer treatment. Over the years, our laboratory studied and characterized in detail the catalytic domain of PDK1, as well as the selective activation of substrates such as SGK or S6K, which in order to be phosphorylated require a docking interaction with a hydrophobic site in PDK1 termed the PIF-Pocket. However, this is not the case of Akt/PKB, since it can be activated in a PIF-Pocket independent way. On the other hand, up to date little is known about the mechanistic and structural details of PDK1 full length. Therefore, we are using an interdisciplinary approach to understand how the full length protein is regulated and how this regulation mechanism can be manipulated to specifically inhibit the activation of PKB/Akt. As a result of a medium-scale screening of small compounds we carried out using AlphaScreen technology, we validated a series of small compounds "hits" that modulate PDK1 structure by interaction at different sites on PDK1. We performed hydrogen/deuterium exchange (HDX) experimentsand crystallization screenings to understand the structure of full length PDK1 and how it can be modulated with small compounds. We here present a series of results obtained using HDX on full length PDK1 and the crystal structure of the catalytic domain of PDK1 bound to a small compound identified in our screening. We also present our in vitro studies to understand the oligomerization of PDK1 visualizing single particles using STORM fluorescence microscopy. Finally, we integrate our data to present an updated model on the molecular mechanism of regulation of PDK1.
publishDate 2019
dc.date.none.fl_str_mv 2019
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/11336/155081
A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1); LV Reunión Científica anual SAIB y XIV congreso de PABMB; Salta; Argentina; 2019; 95-95
0327-9545
CONICET Digital
CONICET
url http://hdl.handle.net/11336/155081
identifier_str_mv A Chemical Biology Approach to Understand The Regulation Of Full-Length Phosphoinositide-Dependent Protein Kinase 1 (Pdk1); LV Reunión Científica anual SAIB y XIV congreso de PABMB; Salta; Argentina; 2019; 95-95
0327-9545
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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