Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors

Detalhes bibliográficos
Autor(a) principal: Felix, J
Data de Publicação: 2019
Outros Autores: Weinhäupl, K, Chipot, C, Dehez, F, Hessel, A, Gauto, DF, Morlot, C, Abian, O, Gutsche, I, Velazquez-Campoy, A, Schanda, P, Fraga, H
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: https://hdl.handle.net/10216/136238
Resumo: Coordinated conformational transitions in oligomeric enzymatic complexes modulate function in response to substrates and play a crucial role in enzyme inhibition and activation. Caseinolytic protease (ClpP) is a tetradecameric complex, which has emerged as a drug target against multiple pathogenic bacteria. Activation of different ClpPs by inhibitors has been independently reported from drug development efforts, but no rationale for inhibitor-induced activation has been hitherto proposed. Using an integrated approach that includes x-ray crystallography, solid- and solution-state nuclear magnetic resonance, molecular dynamics simulations, and isothermal titration calorimetry, we show that the proteasome inhibitor bortezomib binds to the ClpP active-site serine, mimicking a peptide substrate, and induces a concerted allosteric activation of the complex. The bortezomib-activated conformation also exhibits a higher affinity for its cognate unfoldase ClpX. We propose a universal allosteric mechanism, where substrate binding to a single subunit locks ClpP into an active conformation optimized for chaperone association and protein processive degradation.
id RCAP_22b57a01ab2abc91678c1c46d0d18319
oai_identifier_str oai:repositorio-aberto.up.pt:10216/136238
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitorsCoordinated conformational transitions in oligomeric enzymatic complexes modulate function in response to substrates and play a crucial role in enzyme inhibition and activation. Caseinolytic protease (ClpP) is a tetradecameric complex, which has emerged as a drug target against multiple pathogenic bacteria. Activation of different ClpPs by inhibitors has been independently reported from drug development efforts, but no rationale for inhibitor-induced activation has been hitherto proposed. Using an integrated approach that includes x-ray crystallography, solid- and solution-state nuclear magnetic resonance, molecular dynamics simulations, and isothermal titration calorimetry, we show that the proteasome inhibitor bortezomib binds to the ClpP active-site serine, mimicking a peptide substrate, and induces a concerted allosteric activation of the complex. The bortezomib-activated conformation also exhibits a higher affinity for its cognate unfoldase ClpX. We propose a universal allosteric mechanism, where substrate binding to a single subunit locks ClpP into an active conformation optimized for chaperone association and protein processive degradation.American Association for the Advancement of Science20192019-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10216/136238eng2375-254810.1126/sciadv.aaw3818Felix, JWeinhäupl, KChipot, CDehez, FHessel, AGauto, DFMorlot, CAbian, OGutsche, IVelazquez-Campoy, ASchanda, PFraga, Hinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-29T12:45:58Zoai:repositorio-aberto.up.pt:10216/136238Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T23:26:19.136213Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
title Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
spellingShingle Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
Felix, J
title_short Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
title_full Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
title_fullStr Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
title_full_unstemmed Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
title_sort Mechanism of the allosteric activation of the ClpP protease machinery by substrates and active-site inhibitors
author Felix, J
author_facet Felix, J
Weinhäupl, K
Chipot, C
Dehez, F
Hessel, A
Gauto, DF
Morlot, C
Abian, O
Gutsche, I
Velazquez-Campoy, A
Schanda, P
Fraga, H
author_role author
author2 Weinhäupl, K
Chipot, C
Dehez, F
Hessel, A
Gauto, DF
Morlot, C
Abian, O
Gutsche, I
Velazquez-Campoy, A
Schanda, P
Fraga, H
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Felix, J
Weinhäupl, K
Chipot, C
Dehez, F
Hessel, A
Gauto, DF
Morlot, C
Abian, O
Gutsche, I
Velazquez-Campoy, A
Schanda, P
Fraga, H
description Coordinated conformational transitions in oligomeric enzymatic complexes modulate function in response to substrates and play a crucial role in enzyme inhibition and activation. Caseinolytic protease (ClpP) is a tetradecameric complex, which has emerged as a drug target against multiple pathogenic bacteria. Activation of different ClpPs by inhibitors has been independently reported from drug development efforts, but no rationale for inhibitor-induced activation has been hitherto proposed. Using an integrated approach that includes x-ray crystallography, solid- and solution-state nuclear magnetic resonance, molecular dynamics simulations, and isothermal titration calorimetry, we show that the proteasome inhibitor bortezomib binds to the ClpP active-site serine, mimicking a peptide substrate, and induces a concerted allosteric activation of the complex. The bortezomib-activated conformation also exhibits a higher affinity for its cognate unfoldase ClpX. We propose a universal allosteric mechanism, where substrate binding to a single subunit locks ClpP into an active conformation optimized for chaperone association and protein processive degradation.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019-01-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://hdl.handle.net/10216/136238
url https://hdl.handle.net/10216/136238
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 2375-2548
10.1126/sciadv.aaw3818
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Association for the Advancement of Science
publisher.none.fl_str_mv American Association for the Advancement of Science
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
_version_ 1799135569275518976