Structural and dynamic insights revealing how lipase binding domain MD1 of Pseudomonas aeruginosa foldase affects lipase activation
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , , |
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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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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7160 |
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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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1799138044613230592 |