Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis

Alexander M. Walter, Rainer Müller, Bassam Tawfik, Keimpe Db Wierda, Paulo S. Pinheiro, André Nadler, Anthony W. McCarthy, Iwona Ziomkiewicz, Martin Kruse, Gregor Reither, Jens Rettig, Martin Lehmann, Volker Haucke, Bertil Hille, Carsten Schultz, Jakob Balslev Sørensen

Research output: Contribution to journalArticle

11 Citations (Scopus)

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 its 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 may bypass 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.

Original languageEnglish (US)
JournaleLife
Volume6
DOIs
StatePublished - Oct 25 2017
Externally publishedYes

Fingerprint

Exocytosis
Phosphatidylinositols
Fusion reactions
Synaptotagmin I
Lipids
Metabolites
Metabolism
Proteins
Chemical activation
Membranes
Chromaffin Cells
Sensors
Carrier Proteins
Light

Keywords

  • adrenal chromaffin cell
  • cell biology
  • exocytosis
  • mouse
  • Munc13
  • neuroscience
  • optical uncaging
  • phosphatidylinositols
  • synaptotagmin

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

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

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; Sørensen, Jakob Balslev.

In: eLife, Vol. 6, 25.10.2017.

Research output: Contribution to journalArticle

Walter, AM, Müller, R, Tawfik, B, Wierda, KD, Pinheiro, PS, Nadler, A, McCarthy, AW, Ziomkiewicz, I, Kruse, M, Reither, G, Rettig, J, Lehmann, M, Haucke, V, Hille, B, Schultz, C & Sørensen, JB 2017, 'Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis', eLife, vol. 6. https://doi.org/10.7554/eLife.30203
Walter AM, Müller R, Tawfik B, Wierda KD, Pinheiro PS, Nadler A et al. Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis. eLife. 2017 Oct 25;6. https://doi.org/10.7554/eLife.30203
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 ; Sørensen, Jakob Balslev. / Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis. In: eLife. 2017 ; Vol. 6.
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AU - Pinheiro, Paulo S.

AU - Nadler, André

AU - McCarthy, Anthony W.

AU - Ziomkiewicz, Iwona

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AU - Reither, Gregor

AU - Rettig, Jens

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