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Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis. | LitMetric
Phosphatidylinositol-4,5-bisphosphate [PI(4,5)P] is essential for exocytosis. Classical ways of manipulating PI(4,5)P levels are slower than its metabolism, making it difficult to distinguish effects of PI(4,5)P from those of its metabolites. We developed a membrane-permeant, photoactivatable PI(4,5)P, which is loaded into cells in an inactive form and activated by light, allowing sub-second increases in PI(4,5)P levels. By combining this compound with electrophysiological measurements in mouse adrenal chromaffin cells, we show that PI(4,5)P uncaging potentiates exocytosis and identify synaptotagmin-1 (the Ca sensor for exocytosis) and Munc13-2 (a vesicle priming protein) as the relevant effector proteins. PI(4,5)P activation of exocytosis did not depend on the PI(4,5)P-binding CAPS-proteins, suggesting that PI(4,5)P uncaging may bypass CAPS-function. Finally, PI(4,5)P uncaging triggered the rapid fusion of a subset of readily-releasable vesicles, revealing a rapid role of PI(4,5)P in fusion triggering. Thus, optical uncaging of signaling lipids can uncover their rapid effects on cellular processes and identify lipid effectors.
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