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dc.contributor.author
Perez, Camilo
dc.contributor.author
Köhler, Martin
dc.contributor.author
Janser, Daniel
dc.contributor.author
Pardon, Els
dc.contributor.author
Steyaert, Jan
dc.contributor.author
Zenobi, Renato
dc.contributor.author
Locher, Kaspar P.
dc.date.accessioned
2018-10-16T12:27:43Z
dc.date.available
2017-06-12T21:00:48Z
dc.date.available
2018-01-22T15:04:03Z
dc.date.available
2018-01-11T11:56:45Z
dc.date.available
2018-01-22T15:01:42Z
dc.date.available
2018-10-16T12:27:43Z
dc.date.issued
2017-04-19
dc.identifier.issn
2045-2322
dc.identifier.other
10.1038/srep46641
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/233731
dc.identifier.doi
10.3929/ethz-b-000233731
dc.description.abstract
PglK is an ABC transporter that flips a lipid-linked oligosaccharide (LLO) that serves as a donor in protein N-glycosylation. Previous structures revealed two inward-facing conformations, both with very large separations of the nucleotide binding domains (NBDs), and a closed, ADP-bound state that featured an occluded cavity. To investigate additional states, we developed conformation-sensitive, single-domain camelid nanobodies (Nb) and studied their effect on PglK activity. Biochemical, structural, and mass spectrometric analyses revealed that one inhibitory Nb binds as a single copy to homodimeric PglK. The co-crystal structure of this Nb and ADP-bound PglK revealed a new, narrowly inward-open conformation. Rather than inducing asymmetry in the PglK homodimer, the binding of one Nb results in steric constraints that prevent a second Nb to access the symmetry-related site in PglK. The Nb performed its inhibitory role by a “sticky-doorstop” mechanism, where inhibition of ATP hydrolysis and LLO flipping activity occurs due to impaired closing of the NBD interface, which prevents PglK from converting to an outward-open conformation. This inhibitory mode suggests tight conformational coupling between the ATPase sites, which may apply to other ABC transporters.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Nature
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.subject
X-Ray Crystallography
en_US
dc.subject
Biochemistry
en_US
dc.title
Structural basis of inhibition of lipid-linked oligosaccharide flippase PglK by a conformational nanobody
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
dc.date.published
2017-04-19
ethz.journal.title
Scientific Reports
ethz.journal.volume
7
en_US
ethz.journal.abbreviated
Sci Rep
ethz.pages.start
46641
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.identifier.nebis
006751867
ethz.publication.place
London
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02020 - Dep. Chemie und Angewandte Biowiss. / Dep. of Chemistry and Applied Biosc.::02514 - Laboratorium für Organische Chemie / Laboratory of Organic Chemistry::03430 - Zenobi, Renato / Zenobi, Renato
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
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02020 - Dep. Chemie und Angewandte Biowiss. / Dep. of Chemistry and Applied Biosc.::02514 - Laboratorium für Organische Chemie / Laboratory of Organic Chemistry::03430 - Zenobi, Renato / Zenobi, Renato
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
en_US
ethz.grant.agreementno
166672
ethz.grant.fundername
SNF
ethz.grant.funderDoi
10.13039/501100001711
ethz.grant.program
Exzellenzbeitrag in Lebenswissenschaften
ethz.date.deposited
2017-06-12T21:01:11Z
ethz.source
ECIT
ethz.source
FORM
ethz.identifier.importid
imp5936556ed0a7576011
ethz.ecitpid
pub:193814
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2018-01-24T13:46:59Z
ethz.rosetta.lastUpdated
2024-02-02T06:22:27Z
ethz.rosetta.versionExported
true
dc.identifier.olduri
http://hdl.handle.net/20.500.11850/130783
dc.identifier.olduri
http://hdl.handle.net/20.500.11850/227599
ethz.COinS
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