Measuring ligand-cell surface receptor affinities with axial line-scanning fluorescence correlation spectroscopy
release_rev_4d2b6881-681d-44bd-ac81-b53046e3dc8f
by
Antonia Franziska Eckert, Peng Gao, Janine Wesslowski, Xianxian Wang, Jasmijn Rath, Karin Nienhaus, Gary Davidson, Gerd Ulrich Nienhaus
Abstract
Development and homeostasis of multicellular organisms is largely controlled by complex cell-cell signaling networks that rely on specific binding of secreted ligands to cell surface receptors. The Wnt signaling network, as an example, involves multiple ligands and receptors to elicit specific cellular responses. To understand the mechanisms of such a network, ligand-receptor interactions should be characterized quantitatively, ideally in live cells or tissues. Such measurements are possible using fluorescence microscopy yet challenging due to sample movement, low signal-to-background ratio and photobleaching. Here we present a robust approach based on fluorescence correlation spectroscopy with ultra-high speed axial line scanning, yielding precise equilibrium dissociation coefficients of interactions in the Wnt signaling pathway. Using CRISPR/Cas9 editing to endogenously tag receptors with fluorescent proteins, we demonstrate that the method delivers precise results even with low, near-native amounts of receptors.
In application/xml+jats
format
Open Access Publication
In DOAJ
In ISSN ROAD
In Keepers Registry
ISSN-L:
2050-084X
access all versions, variants, and formats of this works (eg, pre-prints)
Crossref Metadata (via API)
Worldcat
SHERPA/RoMEO (journal policies)
wikidata.org
CORE.ac.uk
Semantic Scholar
Google Scholar
This is a specific, static metadata record, not necessarily linked to any current entity in the catalog.