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dc.contributor.author
Napiórkowska, Maja
dc.contributor.author
Boilevin, Jérémy
dc.contributor.author
Darbre, Tamis
dc.contributor.author
Reymond, Jean-Louis
dc.contributor.author
Locher, Kaspar P.
dc.date.accessioned
2018-11-28T13:06:34Z
dc.date.available
2018-11-16T05:43:20Z
dc.date.available
2018-11-28T13:06:34Z
dc.date.issued
2018
dc.identifier.issn
2045-2322
dc.identifier.other
10.1038/s41598-018-34534-0
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/304172
dc.identifier.doi
10.3929/ethz-b-000304172
dc.description.abstract
Oligosaccharyltransferase (OST) is a key enzyme of the N-glycosylation pathway, where it catalyzes the transfer of a glycan from a lipid-linked oligosaccharide (LLO) to an acceptor asparagine within the conserved sequon N-X-T/S. A previous structure of a ternary complex of bacterial single subunit OST, PglB, bound to a non-hydrolyzable LLO analog and a wild type acceptor peptide showed how both substrates bind and how an external loop (EL5) of the enzyme provided specific substrate-binding contacts. However, there was a relatively large separation of the substrates at the active site. Here we present the X-ray structure of PglB bound to a reactive LLO analog and an inhibitory peptide, revealing previously unobserved interactions in the active site. We found that the atoms forming the N-glycosidic bond (C-1 of the GlcNAc moiety of LLO and the –NH2 group of the peptide) are closer than in the previous structure, suggesting that we have captured a conformation closer to the transition state of the reaction. We find that the distance between the divalent metal ion and the glycosidic oxygen of LLO is now 4 Å, suggesting that the metal stabilizes the leaving group of the nucleophilic substitution reaction. Further, the carboxylate group of a conserved aspartate of PglB mediates an interaction network between the reducing-end sugar of the LLO, the asparagine side chain of the acceptor peptide, and a bound divalent metal ion. The interactions identified in this novel state are likely to be relevant in the catalytic mechanisms of all OSTs.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Nature Publishing Group
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
Structure of bacterial oligosaccharyltransferase PglB bound to a reactive LLO and an inhibitory peptide
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
dc.date.published
2018-11-02
ethz.journal.title
Scientific Reports
ethz.journal.volume
8
en_US
ethz.journal.issue
1
en_US
ethz.pages.start
16297
en_US
ethz.size
9 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.grant
Structural and mechanistic studies of components of bacterial protein N-glycosylation pathway and of vitamin B12 transport
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
London
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02521 - Inst. f. Molekularbiologie u. Biophysik / Inst. Molecular Biology and Biophysics::03652 - Locher, Kaspar / Locher, Kaspar
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02521 - Inst. f. Molekularbiologie u. Biophysik / Inst. Molecular Biology and Biophysics::03652 - Locher, Kaspar / Locher, Kaspar
ethz.grant.agreementno
166672
ethz.grant.fundername
SNF
ethz.grant.funderDoi
10.13039/501100001711
ethz.grant.program
ethz.date.deposited
2018-11-16T05:43:21Z
ethz.source
SCOPUS
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2018-11-28T13:06:42Z
ethz.rosetta.lastUpdated
2019-02-03T11:50:06Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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