Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation

Detalhes bibliográficos
Autor(a) principal: Viegas, Aldino
Data de Publicação: 2020
Outros Autores: Dollinger, Peter, Verma, Neha, Kubiak, Jakub, Viennet, Thibault, Seidel, Claus A. M., Gohlke, Holger, Etzkorn, Manuel, Kovacic, Filip, Jaeger, Karl Erich
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/117161
Resumo: ID: HDD16, project IDs: 11237 ET 103/2-1 to ME; no. SE 1195/16-1 no. GO 1367/1-1 JA 448/8-1 ET 103/2-1 PINFRA/22161/2016 -1
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spelling Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activationGeneralID: HDD16, project IDs: 11237 ET 103/2-1 to ME; no. SE 1195/16-1 no. GO 1367/1-1 JA 448/8-1 ET 103/2-1 PINFRA/22161/2016 -1Folding and cellular localization of many proteins of Gram-negative bacteria rely on a network of chaperones and secretion systems. Among them is the lipase-specific foldase Lif, a membrane-bound steric chaperone that tightly binds (KD = 29 nM) and mediates folding of the lipase LipA, a virulence factor of the pathogenic bacterium P. aeruginosa. Lif consists of five-domains, including a mini domain MD1 essential for LipA folding. However, the molecular mechanism of Lif-assisted LipA folding remains elusive. Here, we show in in vitro experiments using a soluble form of Lif (sLif) that isolated MD1 inhibits sLif-assisted LipA activation. Furthermore, the ability to activate LipA is lost in the variant sLifY99A, in which the evolutionary conserved amino acid Y99 from helix α1 of MD1 is mutated to alanine. This coincides with an approximately three-fold reduced affinity of the variant to LipA together with increased flexibility of sLifY99A in the complex as determined by polarization-resolved fluorescence spectroscopy. We have solved the NMR solution structures of P. aeruginosa MD1 and variant MD1Y99A revealing a similar fold indicating that a structural modification is likely not the reason for the impaired activity of variant sLifY99A. Molecular dynamics simulations of the sLif:LipA complex in connection with rigidity analyses suggest a long-range network of interactions spanning from Y99 of sLif to the active site of LipA, which might be essential for LipA activation. These findings provide important details about the putative mechanism for LipA activation and point to a general mechanism of protein folding by multi-domain steric chaperones.UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaRUNViegas, AldinoDollinger, PeterVerma, NehaKubiak, JakubViennet, ThibaultSeidel, Claus A. M.Gohlke, HolgerEtzkorn, ManuelKovacic, FilipJaeger, Karl Erich2021-05-05T23:28:15Z2020-12-012020-12-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/117161eng2045-2322PURE: 29571336https://doi.org/10.1038/s41598-020-60093-4info: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:RCAAP2024-03-11T05:00:13Zoai:run.unl.pt:10362/117161Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:43:30.746319Repositório 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 Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
title Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
spellingShingle Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
Viegas, Aldino
General
title_short Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
title_full Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
title_fullStr Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
title_full_unstemmed Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
title_sort Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
author Viegas, Aldino
author_facet Viegas, Aldino
Dollinger, Peter
Verma, Neha
Kubiak, Jakub
Viennet, Thibault
Seidel, Claus A. M.
Gohlke, Holger
Etzkorn, Manuel
Kovacic, Filip
Jaeger, Karl Erich
author_role author
author2 Dollinger, Peter
Verma, Neha
Kubiak, Jakub
Viennet, Thibault
Seidel, Claus A. M.
Gohlke, Holger
Etzkorn, Manuel
Kovacic, Filip
Jaeger, Karl Erich
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv UCIBIO - Applied Molecular Biosciences Unit
DQ - Departamento de Química
RUN
dc.contributor.author.fl_str_mv Viegas, Aldino
Dollinger, Peter
Verma, Neha
Kubiak, Jakub
Viennet, Thibault
Seidel, Claus A. M.
Gohlke, Holger
Etzkorn, Manuel
Kovacic, Filip
Jaeger, Karl Erich
dc.subject.por.fl_str_mv General
topic General
description ID: HDD16, project IDs: 11237 ET 103/2-1 to ME; no. SE 1195/16-1 no. GO 1367/1-1 JA 448/8-1 ET 103/2-1 PINFRA/22161/2016 -1
publishDate 2020
dc.date.none.fl_str_mv 2020-12-01
2020-12-01T00:00:00Z
2021-05-05T23:28:15Z
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 http://hdl.handle.net/10362/117161
url http://hdl.handle.net/10362/117161
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 2045-2322
PURE: 29571336
https://doi.org/10.1038/s41598-020-60093-4
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.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
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