Measuring ligand-cell surface receptor affinities with axial line-scanning fluorescence correlation spectroscopy release_vskjrnpnhfc6zc4gzh4ibvwzme [as of editgroup_3ayixglgizamtlrqopwk33mfvi]

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.
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Published in eLife by eLife Sciences Publications, Ltd
ISSN-L 2050-084X
Volume 9
Release Date 2020-05-22
Publisher eLife Sciences Publications, Ltd
Primary Language en (lookup)
Type  article-journal
Stage   published
Date   2020-05-22
DOI  10.7554/elife.55286
PubMed  32441251
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Open Access Publication
In DOAJ
In ISSN ROAD
ISSN-L:  2050-084X
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