Annexin-chromaffin granule membrane interactions: a comparative study of synexin, p32 and p67.
The chromaffin granule membrane binding and aggregating properties of three annexins, synexin, p32 and p67, have been studied and compared. Each protein was activated to bind and aggregate membranes with a biphasic Ca2+ dependence, with one phase titrating between pCa 5.0-3.5 and the second at higher levels of calcium (pCa less than 3.5). cis-Unsaturated free fatty acids lowered these Ca2+ requirements by approximately one log unit. Barium and strontium were able to partially substitute for calcium, with the order of sensitivity Ca2+ greater than Sr2+ greater than Ba2+. The proteins appeared to bind to distinct but overlapping populations of receptor sites, and did so in a manner displaying positive cooperativity at the higher Ca2+ levels. The maximal efficacy of the proteins as membrane aggregators differed with synexin being 1-2-fold more efficacious than p32, which in turn was 7-fold more efficacious than p67. In combination, p67 was an effective inhibitor of granule aggregation induced by synexin or p32, while p32 was able to both promote and inhibit synexin-induced granule aggregation in a manner which varied with synexin concentration. The complexity of these annexin-membrane interactions may be a reflection of the multidomain structure of the annexins and may have implications for the differential functions of these proteins in cells.
Zaks
W J
WJ
Department of Pharmacology, University of Virginia, Charlottesville 22908.
Creutz
C E
CE
D000818:Animals; D017310:Annexin A7; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002837:Chromaffin Granules / Q000187:drug effects / Q000378:metabolism*; D005227:Fatty Acids / Q000494:pharmacology; D008099:Liver / Q000737:chemistry; D011506:Proteins / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D038981:Receptors, Collagen
D017310:Annexin A7; D002135:Calcium-Binding Proteins; D005227:Fatty Acids; D011506:Proteins; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; D002118:Calcium