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dc.contributor.author
Klose, Daniel
dc.contributor.author
Holla, Andrea
dc.contributor.author
Gmeiner, Christoph
dc.contributor.author
Nettels, Daniel
dc.contributor.author
Ritsch, Irina
dc.contributor.author
Bross, Nadja
dc.contributor.author
Yulikov, Maxim
dc.contributor.author
Allain, Frédéric H.-T.
dc.contributor.author
Schuler, Benjamin
dc.contributor.author
Jeschke, Gunnar
dc.date.accessioned
2021-11-04T15:45:36Z
dc.date.available
2021-09-29T09:31:42Z
dc.date.available
2021-09-29T10:01:25Z
dc.date.available
2021-11-04T15:45:36Z
dc.date.issued
2021-11-02
dc.identifier.issn
0006-3495
dc.identifier.issn
1542-0086
dc.identifier.other
10.1016/j.bpj.2021.09.021
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/507587
dc.identifier.doi
10.3929/ethz-b-000507587
dc.description.abstract
Förster resonance energy transfer (FRET) and electron paramagnetic resonance (EPR) spectroscopy are complementary techniques for quantifying distances in the nanometer range. Both approaches are commonly employed for probing the conformations and conformational changes of biological macromolecules based on site-directed fluorescent or paramagnetic labeling. FRET can be applied in solution at ambient temperature and thus provides direct access to dynamics, especially if used at the single-molecule level, whereas EPR requires immobilization or work at cryogenic temperatures but provides data that can be more reliably used to extract distance distributions. However, a combined analysis of the complementary data from the two techniques has been complicated by the lack of a common modeling framework. Here, we demonstrate a systematic analysis approach based on rotamer libraries for both FRET and EPR labels to predict distance distributions between two labels from a structural model. Dynamics of the fluorophores within these distance distributions are taken into account by diffusional averaging, which improves the agreement with experiment. Benchmarking this methodology with a series of surface-exposed pairs of sites in a structured protein domain reveals that the lowest resolved distance differences can be as small as ∼0.25 nm for both techniques, with quantitative agreement between experimental and simulated transfer efficiencies within a range of ±0.045. Rotamer library analysis thus establishes a coherent way of treating experimental data from EPR and FRET and provides a basis for integrative structural modeling, including studies of conformational distributions and dynamics of biological macromolecules using both techniques.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Cell Press
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
Resolving distance variations by single-molecule FRET and EPR spectroscopy using rotamer libraries
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
dc.date.published
2021-09-16
ethz.journal.title
Biophysical Journal
ethz.journal.volume
120
en_US
ethz.journal.issue
21
en_US
ethz.journal.abbreviated
Biophys. j.
ethz.pages.start
4842
en_US
ethz.pages.end
4858
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.grant
Role of Disordered Regions in RNA-Binding Proteins for Function and Pathology
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
Bethesda, MD
en_US
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.::02515 - Laboratorium für Physikalische Chemie / Laboratory of Physical Chemistry::03810 - Jeschke, Gunnar / Jeschke, Gunnar
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02517 - Institut für Biochemie / Institute of Biochemistry (IBC)::03591 - Allain, Frédéric / Allain, Frédéric
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.::02515 - Laboratorium für Physikalische Chemie / Laboratory of Physical Chemistry::03810 - Jeschke, Gunnar / Jeschke, Gunnar
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02517 - Institut für Biochemie / Institute of Biochemistry (IBC)::03591 - Allain, Frédéric / Allain, Frédéric
ethz.grant.agreementno
170976
ethz.grant.fundername
SNF
ethz.grant.funderDoi
10.13039/501100001711
ethz.grant.program
Sinergia
ethz.date.deposited
2021-09-29T09:31:47Z
ethz.source
FORM
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2021-11-04T15:45:48Z
ethz.rosetta.lastUpdated
2021-11-04T15:45:48Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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