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

2018-12-01Artigo científico Acesso aberto
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dc.contributor.authorLeisico, Francisco
dc.contributor.authorVieira, Diana V.
dc.contributor.authorFigueiredo, Teresa A.
dc.contributor.authorSilva, Micael
dc.contributor.authorCabrita, Eurico J.
dc.contributor.authorSobral, Rita G.
dc.contributor.authorLudovice, Ana Madalena
dc.contributor.authorTrincão, José
dc.contributor.authorRomão, Maria João
dc.contributor.authorDe Lencastre, Hermínia
dc.contributor.authorSantos-Silva, Teresa
dc.contributor.institutionUCIBIO - Applied Molecular Biosciences Unit
dc.contributor.institutionDQ - Departamento de Química
dc.contributor.institutionInstituto de Tecnologia Química e Biológica António Xavier (ITQB)
dc.contributor.institutionDCV - Departamento de Ciências da Vida
dc.contributor.institutionMolecular, Structural and Cellular Microbiology (MOSTMICRO)
dc.contributor.pblNature Publishing Group
dc.date.accessioned2019-01-25T23:38:16Z
dc.date.available2019-01-25T23:38:16Z
dc.date.issued2018-12-01
dc.descriptionThe 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).
dc.description.abstractGram-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.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent2522024
dc.identifier.doi10.1038/s41598-018-22986-3
dc.identifier.issn2045-2322
dc.identifier.otherPURE: 3842359
dc.identifier.otherPURE UUID: 2f333d5c-c4da-48f2-935c-258e79f7a786
dc.identifier.otherScopus: 85044530342
dc.identifier.otherWOS: 000428518900010
dc.identifier.otherPubMed: 29593310
dc.identifier.otherPubMedCentral: PMC5871853
dc.identifier.otherORCID: /0000-0002-3004-0543/work/54825303
dc.identifier.otherORCID: /0000-0003-4533-7531/work/201987283
dc.identifier.urihttp://www.scopus.com/inward/record.url?scp=85044530342&partnerID=8YFLogxK
dc.identifier.urlhttps://www.scopus.com/pages/publications/85044530342
dc.language.isoeng
dc.peerreviewedyes
dc.relationinfo:eu-repo/grantAgreement/FCT/5876/147258/PT
dc.subjectArginine
dc.subjectLigases
dc.subjectArginine biosynthesis
dc.subjectGeneral
dc.titleFirst insights of peptidoglycan amidation in Gram-positive bacteria-The high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatDen
dc.typejournal article
degois.publication.issue1
degois.publication.titleScientific Reports
degois.publication.volume8
dspace.entity.typePublication
oaire.awardNumberUID/Multi/04378/2013
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FMulti%2F04378%2F2013/PT
oaire.fundingStream5876
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccess
relation.isProjectOfPublicationf6c6102f-c577-4633-a70c-08a0b27e1207
relation.isProjectOfPublication.latestForDiscoveryf6c6102f-c577-4633-a70c-08a0b27e1207

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