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MBC in Press, published online ahead of print January 5, 2005
Mol. Biol. Cell 10.1091/mbc.E04-07-0627

A more recent version of this article appeared on March 1, 2005
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Submitted on July 23, 2004
Accepted on December 16, 2004

Annexin 2 Promotes the Formation of Lipid Micro-domains Required for Calcium-regulated Exocytosis of Dense-core Vesicles

Sylvette Chasserot-Golaz,*{dagger} Nicolas Vitale,* Emeline Umbrecht-Jenck,* Derek Knight,{ddagger} Volker Gerke,{sect} and Marie-France Bader*

*Centre National de la Recherche Scientifique, Unité Propre de Recherche 2356, IFR 37 des Neurosciences, 67084 Strasbourg Cedex, France; {ddagger}Biomedical Sciences, Kings College London, London SE1 1U, United Kingdom; {sect}Institute for Medical Biochemistry, ZMBE, University of Münster, D-48149 Münster, Germany

Monitoring Editor: Anthony Bretscher

Annexin 2 is a calcium-dependent phospholipid-binding protein that has been implicated in a number of membrane-related 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 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 was also 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.

{dagger}Corresponding author. E-mail: chasserot{at}neurochem.u-strasbg.fr






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