dc.contributor.author
Kashyap, Purba
dc.contributor.author
Bertelli, Sara
dc.contributor.author
Cao, Fakun
dc.contributor.author
Kostritskaia, Yulia
dc.contributor.author
Blank, Fenja
dc.contributor.author
Srikanth, Niranjan A.
dc.contributor.author
Schlack-Leigers, Claire
dc.contributor.author
Saleppico, Roberto
dc.contributor.author
Bierhuizen, Dolf
dc.contributor.author
Ewers, Helge
dc.date.accessioned
2024-04-16T09:50:31Z
dc.date.available
2024-04-16T09:50:31Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/42721
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-42440
dc.description.abstract
We developed a system for optogenetic release of single molecules in cells. We confined soluble and transmembrane proteins to the Golgi apparatus via a photocleavable protein and released them by short pulses of light. Our method allows for a light dose-dependent delivery of functional proteins to the cytosol and plasma membrane in amounts compatible with single-molecule imaging, greatly simplifying access to single-molecule microscopy of any protein in live cells. We were able to reconstitute ion conductance by delivering BK and LRRC8/volume-regulated anion channels to the plasma membrane. Finally we were able to induce NF-kB signaling in T lymphoblasts stimulated by interleukin-1 by controlled release of a signaling protein that had been knocked out. We observed light-induced formation of functional inflammatory signaling complexes that triggered phosphorylation of the inhibitor of nuclear factor kappa-B kinase only in activated cells. We thus developed an optogenetic method for the reconstitution and investigation of cellular function at the single-molecule level.
en
dc.format.extent
15 Seiten
dc.rights.uri
https://creativecommons.org/licenses/by/4.0/
dc.subject
Optogenetics
en
dc.subject
Single-molecule biophysics
en
dc.subject
Super-resolution microscopy
en
dc.subject.ddc
500 Naturwissenschaften und Mathematik::540 Chemie::540 Chemie und zugeordnete Wissenschaften
dc.title
An optogenetic method for the controlled release of single molecules
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.doi
10.1038/s41592-024-02204-x
dcterms.bibliographicCitation.journaltitle
Nature Methods
dcterms.bibliographicCitation.number
4
dcterms.bibliographicCitation.pagestart
666
dcterms.bibliographicCitation.pageend
672
dcterms.bibliographicCitation.volume
21
dcterms.bibliographicCitation.url
https://doi.org/10.1038/s41592-024-02204-x
refubium.affiliation
Biologie, Chemie, Pharmazie
refubium.affiliation.other
Institut für Chemie und Biochemie
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refubium.funding
Springer Nature DEAL
refubium.note.author
Die Publikation wurde aus Open Access Publikationsgeldern der Freien Universität Berlin gefördert.
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.isPartOf.eissn
1548-7105