Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis
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Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis. / Walter, Alexander M; Müller, Rainer; Tawfik, Bassam; Wierda, Keimpe Db; Pinheiro, Paulo S; Nadler, André; McCarthy, Anthony W; Ziomkiewicz, Iwona; Kruse, Martin; Reither, Gregor; Rettig, Jens; Lehmann, Martin; Haucke, Volker; Hille, Bertil; Schultz, Carsten; Sorensen, Jakob Balslev.
In: eLife, Vol. 6, e30203, 2017.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis
AU - Walter, Alexander M
AU - Müller, Rainer
AU - Tawfik, Bassam
AU - Wierda, Keimpe Db
AU - Pinheiro, Paulo S
AU - Nadler, André
AU - McCarthy, Anthony W
AU - Ziomkiewicz, Iwona
AU - Kruse, Martin
AU - Reither, Gregor
AU - Rettig, Jens
AU - Lehmann, Martin
AU - Haucke, Volker
AU - Hille, Bertil
AU - Schultz, Carsten
AU - Sorensen, Jakob Balslev
PY - 2017
Y1 - 2017
N2 - Phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] is essential for exocytosis. Classical ways of manipulating PI(4,5)P2 levels are slower than metabolism, making it difficult to distinguish effects of PI(4,5)P2 from those of its metabolites. We developed a membrane-permeant, photoactivatable PI(4,5)P2, which is loaded into cells in an inactive form and activated by light, allowing sub-second increases in PI(4,5)P2 levels. By combining this compound with electrophysiological measurements in mouse adrenal chromaffin cells, we show that PI(4,5)P2 uncaging potentiates exocytosis and identify synaptotagmin-1 (the Ca(2+) sensor for exocytosis) and Munc13-2 (a vesicle priming protein) as the relevant effector proteins. PI(4,5)P2 activation of exocytosis did not depend on the PI(4,5)P2-binding CAPS-proteins, suggesting that PI(4,5)P2 uncaging bypasses CAPS-function. Finally, PI(4,5)P2 uncaging triggered the rapid fusion of a subset of readily-releasable vesicles, revealing a rapid role of PI(4,5)P2 in fusion triggering. Thus, optical uncaging of signaling lipids can uncover their rapid effects on cellular processes and identify lipid effectors.
AB - Phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] is essential for exocytosis. Classical ways of manipulating PI(4,5)P2 levels are slower than metabolism, making it difficult to distinguish effects of PI(4,5)P2 from those of its metabolites. We developed a membrane-permeant, photoactivatable PI(4,5)P2, which is loaded into cells in an inactive form and activated by light, allowing sub-second increases in PI(4,5)P2 levels. By combining this compound with electrophysiological measurements in mouse adrenal chromaffin cells, we show that PI(4,5)P2 uncaging potentiates exocytosis and identify synaptotagmin-1 (the Ca(2+) sensor for exocytosis) and Munc13-2 (a vesicle priming protein) as the relevant effector proteins. PI(4,5)P2 activation of exocytosis did not depend on the PI(4,5)P2-binding CAPS-proteins, suggesting that PI(4,5)P2 uncaging bypasses CAPS-function. Finally, PI(4,5)P2 uncaging triggered the rapid fusion of a subset of readily-releasable vesicles, revealing a rapid role of PI(4,5)P2 in fusion triggering. Thus, optical uncaging of signaling lipids can uncover their rapid effects on cellular processes and identify lipid effectors.
KW - Journal Article
U2 - 10.7554/eLife.30203
DO - 10.7554/eLife.30203
M3 - Journal article
C2 - 29068313
VL - 6
JO - eLife
JF - eLife
SN - 2050-084X
M1 - e30203
ER -
ID: 185030190