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

doi:10.7554/eLife.30203

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Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis.

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Walter, A. M., Müller, R., Tawfik, B., Wierda, K. D., Pinheiro, P. S., Nadler, A., et al. (2017). Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis. eLife, 6: e30203. doi:10.7554/eLife.30203.

Cite as: https://hdl.handle.net/21.11116/0000-0002-8C90-D

Abstract

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 Ca2+ 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.