Exclusive photorelease of signalling lipids at the plasma membrane
Nat Commun. 2015-12-01; 6(1):
Lire sur PubMed
1. Nat Commun. 2015 Dec 21;6:10056. doi: 10.1038/ncomms10056.
Exclusive photorelease of signalling lipids at the plasma membrane.
Nadler A(1)(2), Yushchenko DA(1)(3), Müller R(1), Stein F(1), Feng S(1), Mulle
C(4), Carta M(4), Schultz C(1).
(1)European Molecular Biology Laboratory, Cell Biology and Biophysics Unit,
Meyerhofstraße 1, 69117 Heidelberg, Germany.
(2)Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstraße
108, 01307 Dresden, Germany.
(3)Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the
Czech Republic, Flemingovo náměstí 2, 16610 Prague 6, Czech Republic.
(4)Institut Interdisciplinaire de Neurosciences, CNRS UMR 5297 Université
Bordeaux 2, 146, rue Léo-Saignat, 33077 Bordeaux, France.
Photoactivation of caged biomolecules has become a powerful approach to study
cellular signalling events. Here we report a method for anchoring and uncaging
biomolecules exclusively at the outer leaflet of the plasma membrane by employing
a photocleavable, sulfonated coumarin derivative. The novel caging group allows
quantifying the reaction progress and efficiency of uncaging reactions in a
live-cell microscopy setup, thereby greatly improving the control of uncaging
experiments. We synthesized arachidonic acid derivatives bearing the new
negatively charged or a neutral, membrane-permeant coumarin caging group to
locally induce signalling either at the plasma membrane or on internal membranes
in β-cells and brain slices derived from C57B1/6 mice. Uncaging at the plasma
membrane triggers a strong enhancement of calcium oscillations in β-cells and a
pronounced potentiation of synaptic transmission while uncaging inside cells
blocks calcium oscillations in β-cells and causes a more transient effect on
neuronal transmission, respectively. The precise subcellular site of arachidonic
acid release is therefore crucial for signalling outcome in two independent
PMID: 26686736 [Indexed for MEDLINE]