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Vol. 16, Issue 3, 1108-1119, March 2005

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Annexin 2 Promotes the Formation of Lipid Microdomains Required for Calcium-regulated Exocytosis of Dense-Core Vesicles

Sylvette Chasserot-Golaz ^*, Nicolas Vitale ^*, Emeline Umbrecht-Jenck ^*, Derek Knight , Volker Gerke , and Marie-France Bader ^*

^* Centre National de la Recherche Scientifique, Unité Propre de Recherche 2356, IFR 37 des Neurosciences, 67084 Strasbourg Cedex, France; Biomedical Sciences, Kings College London, London SE1 1U, United Kingdom; and Institute for Medical Biochemistry, Center for Molecular Biology of Inflammation, University of Münster, D-48149 Münster, Germany

Submitted July 23, 2004; Revised November 24, 2004; Accepted December 16, 2004
Monitoring Editor: Anthony Bretscher

Annexin 2 is a calcium-dependent phospholipid-binding protein that has been implicated in a number of membranerelated events, including regulated exocytosis. In chromaffin cells, we previously reported that catecholamine secretion requires the translocation and formation of the annexin 2 tetramer near the exocytotic sites. Here, to obtain direct evidence for a role of annexin 2 in exocytosis, we modified its expression level in chromaffin cells by using the Semliki Forest virus expression system. Using a real-time assay for individual cells, we found that the reduction of cytosolic annexin 2, and the consequent decrease of annexin 2 tetramer at the cell periphery, strongly inhibited exocytosis, most likely at an early stage before membrane fusion. Secretion also was severely impaired in cells expressing a chimera that sequestered annexin 2 into cytosolic aggregates. Moreover, we demonstrate that secretagogue-evoked stimulation triggers the formation of lipid rafts in the plasma membrane, essential for exocytosis, and which can be attributed to the annexin 2 tetramer. We propose that annexin 2 acts as a calcium-dependent promoter of lipid microdomains required for structural and spatial organization of the exocytotic machinery.

Address correspondence to: Sylvette Chasserot-Golaz (chasserot{at}neurochem.u-strasbg.fr).

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