Ca(2+)-dependent annexin self-association on membrane surfaces.
Annexin self-association was studied with 90 degrees light scattering and resonance energy transfer between fluorescein (donor) and eosin (acceptor) labeled proteins. Synexin (annexin VII), p32 (annexin IV), and p67 (annexin VI) self-associated in a Ca(2+)-dependent manner in solution. However, this activity was quite labile and, especially for p32 and p67, was not consistently observed. When bound to chromaffin granule membranes, the three proteins consistently self-associated and did so at Ca2+ levels (pCa 5.0-4.5) approximately 10-fold lower than required when in solution. Phospholipid vesicles containing phosphatidylserine and phosphatidylethanolamine (1:1 or 1:3) were less effective at supporting annexin polymerization than were those containing phosphatidylserine and phosphatidylcholine (1:0, 1:1, or 1:3). The annexins bound chromaffin granule membranes in a positively cooperative manner under conditions where annexin self-association was observed, and both phenomena were inhibited by trifluoperazine. Ca(2+)-dependent chromaffin granule membrane aggregation, induced by p32 or synexin, was associated with intermembrane annexin polymerization at Ca2+ levels less than pCa 4, but not at higher Ca2+ concentrations, suggesting that annexin self-association may be necessary for membrane contact at low Ca2+ levels but not at higher Ca2+ levels where the protein may bind two membranes as a monomer.
Zaks
W J
WJ
Department of Pharmacology, University of Virginia, Charlottesville 22908.
Creutz
C E
CE
D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003432:Cross-Linking Reagents; D004735:Energy Transfer; D008051:Lipid Bilayers / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry; D014268:Trifluoperazine / Q000494:pharmacology
D002135:Calcium-Binding Proteins; D003432:Cross-Linking Reagents; D008051:Lipid Bilayers; D008563:Membrane Lipids; D010743:Phospholipids; D014268:Trifluoperazine; D002118:Calcium