Cholesterol regulates membrane binding and aggregation by annexin 2 at submicromolar Ca(2+) concentration.
Annexin 2 is a member of the annexin family which has been implicated in calcium-regulated exocytosis. This contention is largely based on Ca(2+)-dependent binding of the protein to anionic phospholipids. However, annexin 2 was shown to be associated with chromaffin granules in the presence of EGTA. A fraction of this bound annexin 2 was released by methyl-beta-cyclodextrin, a reagent which depletes cholesterol from membranes. Restoration of the cholesterol content of chromaffin granule membranes with cholesterol/methyl-beta-cyclodextrin complexes restored the Ca(2+)-independent binding of annexin 2. The binding of both, monomeric and tetrameric forms of annexin 2 was also tested on liposomes of different composition. In the absence of Ca(2+), annexin 2, especially in its tetrameric form, bound to liposomes containing phosphatidylserine, and the addition of cholesterol to these liposomes increased the binding. Consistent with this observation, liposomes containing phosphatidylserine and cholesterol were aggregated by the tetrameric form of annexin 2 at submicromolar Ca(2+) concentrations. These results indicate that the lipid composition of membranes, and especially their cholesterol content, is important in the control of the subcellular localization of annexin 2 in resting cells, at low Ca(2+) concentration. Annexin 2 might be associated with membrane domains enriched in phosphatidylserine and cholesterol.
Ayala-Sanmartin
J
J
Unité de Biologie Cellulaire et Moléculaire de la Sécrétion, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, 75005 Paris, France. ayala@ibpc.fr
Henry
J P
JP
Pradel
L A
LA
D017306:Annexin A2 / Q000737:chemistry / Q000378:metabolism*; D002118:Calcium / Q000032:analysis* / Q000494:pharmacology; D002784:Cholesterol / Q000494:pharmacology*; D002837:Chromaffin Granules / Q000378:metabolism; D003505:Cyclodextrins / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D007425:Intracellular Membranes / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry; D008563:Membrane Lipids / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D047392:beta-Cyclodextrins
D017306:Annexin A2; D003505:Cyclodextrins; D008081:Liposomes; D008563:Membrane Lipids; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D002784:Cholesterol; D002118:Calcium