First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD

Detalhes bibliográficos
Autor(a) principal: Leisico, Francisco
Data de Publicação: 2018
Outros Autores: Vieira, Diana V., Figueiredo, Teresa A., Silva, Micael, Cabrita, Eurico J., Sobral, Rita G., Ludovice, Ana Madalena, Trincão, José, Romão, Maria João, De Lencastre, Hermínia, Santos-Silva, Teresa
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: https://doi.org/10.1038/s41598-018-22986-3
Resumo: The authors thank the Oxford Protein Production Facility, Research Complex (Harwell, Didcot, UK) for access to high-throughput facilities and assistance during the experiments; beamline staff at I02 and I04 for assistance during data collection at Diamond Light Source (Didcot, UK). DVV, TAF, FL and MS were supported by fellowships SFRH/BD/62415/2009, SFRH/BD/36843/2007, info:eu-repo/grantAgreement/FCT/5876/147258/PT PD/BD/105737/2014 and PD/BD/128202/2016 from Fundacao para a Ciencia e Tecnologia (FCT-MCTES), Portugal. This project was supported by Project PTDC/BIA-MIC/3195/2012 from FCT-MCTES, Portugal, Project LISBOA-01-0145-FEDER-007660 (Microbiologia Molecular, Estrutural e Celular) funded by FEDER - COMPETE2020 - Programa Operacional Competitividade e Internacionalizacao (POCI), Pest-OE/BIA/UI0457/2001 (CREM) and Unidade de Ciencias Biomoleculares Aplicadas-UCIBIO which is financed by national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728). The NMR spectrometers are part of the National NMR Network (PTNMR) and are supported by Infrastructure Project No 022161 (co-financed by FEDER through COMPETE 2020, POCI, and PORL and FCT through PIDDAC).
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spelling First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatDArginineLigasesArginine biosynthesisGeneralThe authors thank the Oxford Protein Production Facility, Research Complex (Harwell, Didcot, UK) for access to high-throughput facilities and assistance during the experiments; beamline staff at I02 and I04 for assistance during data collection at Diamond Light Source (Didcot, UK). DVV, TAF, FL and MS were supported by fellowships SFRH/BD/62415/2009, SFRH/BD/36843/2007, info:eu-repo/grantAgreement/FCT/5876/147258/PT PD/BD/105737/2014 and PD/BD/128202/2016 from Fundacao para a Ciencia e Tecnologia (FCT-MCTES), Portugal. This project was supported by Project PTDC/BIA-MIC/3195/2012 from FCT-MCTES, Portugal, Project LISBOA-01-0145-FEDER-007660 (Microbiologia Molecular, Estrutural e Celular) funded by FEDER - COMPETE2020 - Programa Operacional Competitividade e Internacionalizacao (POCI), Pest-OE/BIA/UI0457/2001 (CREM) and Unidade de Ciencias Biomoleculares Aplicadas-UCIBIO which is financed by national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728). The NMR spectrometers are part of the National NMR Network (PTNMR) and are supported by Infrastructure Project No 022161 (co-financed by FEDER through COMPETE 2020, POCI, and PORL and FCT through PIDDAC).Gram-positive bacteria homeostasis and antibiotic resistance mechanisms are dependent on the intricate architecture of the cell wall, where amidated peptidoglycan plays an important role. The amidation reaction is carried out by the bi-enzymatic complex MurT-GatD, for which biochemical and structural information is very scarce. In this work, we report the first crystal structure of the glutamine amidotransferase member of this complex, GatD from Staphylococcus aureus, at 1.85 Å resolution. A glutamine molecule is found close to the active site funnel, hydrogen-bonded to the conserved R128. In vitro functional studies using 1H-NMR spectroscopy showed that S. aureus MurT-GatD complex has glutaminase activity even in the absence of lipid II, the MurT substrate. In addition, we produced R128A, C94A and H189A mutants, which were totally inactive for glutamine deamidation, revealing their essential role in substrate sequestration and catalytic reaction. GatD from S. aureus and other pathogenic bacteria share high identity to enzymes involved in cobalamin biosynthesis, which can be grouped in a new sub-family of glutamine amidotransferases. Given the ubiquitous presence of GatD, these results provide significant insights into the molecular basis of the so far undisclosed amidation mechanism, contributing to the development of alternative therapeutics to fight infections.UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaInstituto de Tecnologia Química e Biológica António Xavier (ITQB)DCV - Departamento de Ciências da VidaMolecular, Structural and Cellular Microbiology (MOSTMICRO)RUNLeisico, FranciscoVieira, Diana V.Figueiredo, Teresa A.Silva, MicaelCabrita, Eurico J.Sobral, Rita G.Ludovice, Ana MadalenaTrincão, JoséRomão, Maria JoãoDe Lencastre, HermíniaSantos-Silva, Teresa2019-01-25T23:38:16Z2018-12-012018-12-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://doi.org/10.1038/s41598-018-22986-3eng2045-2322PURE: 3842359http://www.scopus.com/inward/record.url?scp=85044530342&partnerID=8YFLogxKhttps://doi.org/10.1038/s41598-018-22986-3info: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:RCAAP2024-03-11T04:28:17Zoai:run.unl.pt:10362/58648Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:33:17.942629Repositório 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 First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
title First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
spellingShingle First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
Leisico, Francisco
Arginine
Ligases
Arginine biosynthesis
General
title_short First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
title_full First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
title_fullStr First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
title_full_unstemmed First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
title_sort First insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
author Leisico, Francisco
author_facet Leisico, Francisco
Vieira, Diana V.
Figueiredo, Teresa A.
Silva, Micael
Cabrita, Eurico J.
Sobral, Rita G.
Ludovice, Ana Madalena
Trincão, José
Romão, Maria João
De Lencastre, Hermínia
Santos-Silva, Teresa
author_role author
author2 Vieira, Diana V.
Figueiredo, Teresa A.
Silva, Micael
Cabrita, Eurico J.
Sobral, Rita G.
Ludovice, Ana Madalena
Trincão, José
Romão, Maria João
De Lencastre, Hermínia
Santos-Silva, Teresa
author2_role author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv UCIBIO - Applied Molecular Biosciences Unit
DQ - Departamento de Química
Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
DCV - Departamento de Ciências da Vida
Molecular, Structural and Cellular Microbiology (MOSTMICRO)
RUN
dc.contributor.author.fl_str_mv Leisico, Francisco
Vieira, Diana V.
Figueiredo, Teresa A.
Silva, Micael
Cabrita, Eurico J.
Sobral, Rita G.
Ludovice, Ana Madalena
Trincão, José
Romão, Maria João
De Lencastre, Hermínia
Santos-Silva, Teresa
dc.subject.por.fl_str_mv Arginine
Ligases
Arginine biosynthesis
General
topic Arginine
Ligases
Arginine biosynthesis
General
description The authors thank the Oxford Protein Production Facility, Research Complex (Harwell, Didcot, UK) for access to high-throughput facilities and assistance during the experiments; beamline staff at I02 and I04 for assistance during data collection at Diamond Light Source (Didcot, UK). DVV, TAF, FL and MS were supported by fellowships SFRH/BD/62415/2009, SFRH/BD/36843/2007, info:eu-repo/grantAgreement/FCT/5876/147258/PT PD/BD/105737/2014 and PD/BD/128202/2016 from Fundacao para a Ciencia e Tecnologia (FCT-MCTES), Portugal. This project was supported by Project PTDC/BIA-MIC/3195/2012 from FCT-MCTES, Portugal, Project LISBOA-01-0145-FEDER-007660 (Microbiologia Molecular, Estrutural e Celular) funded by FEDER - COMPETE2020 - Programa Operacional Competitividade e Internacionalizacao (POCI), Pest-OE/BIA/UI0457/2001 (CREM) and Unidade de Ciencias Biomoleculares Aplicadas-UCIBIO which is financed by national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728). The NMR spectrometers are part of the National NMR Network (PTNMR) and are supported by Infrastructure Project No 022161 (co-financed by FEDER through COMPETE 2020, POCI, and PORL and FCT through PIDDAC).
publishDate 2018
dc.date.none.fl_str_mv 2018-12-01
2018-12-01T00:00:00Z
2019-01-25T23:38:16Z
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://doi.org/10.1038/s41598-018-22986-3
url https://doi.org/10.1038/s41598-018-22986-3
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 2045-2322
PURE: 3842359
http://www.scopus.com/inward/record.url?scp=85044530342&partnerID=8YFLogxK
https://doi.org/10.1038/s41598-018-22986-3
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)
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instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
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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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