Characterization of the chromobindins. Soluble proteins that bind to the chromaffin granule membrane in the presence of Ca2+.
A group of proteins that bind to the chromaffin granule membrane in the presence of Ca2+ has been isolated by affinity chromatography of bovine adrenal medullary cytosol on granule membranes coupled to Sepharose 4B. Twenty-two of these proteins were resolved into classes depending upon the Ca2+ concentration at which they were eluted from the affinity column (40 or 0.1 microM), upon their affinities for native granule membranes or for liposomes prepared from extracted granule lipids, and upon the requirement of seven of the proteins for ATP in the cytosol fraction and column buffers to promote binding. The molecular weights and isoelectric points of these proteins were determined by two-dimensional electrophoresis. Two of the granule-binding proteins were identified: synexin and calmodulin. Calmodulin was found to bind to seven specific granule membrane proteins after diffusion of 125I-labeled calmodulin into an acrylamide gel of membrane proteins separated by electrophoresis in the presence of sodium dodecyl sulfate. A phospholipid-activated protein kinase activity, possibly due to protein kinase C, was present in the granule-binding fraction. Two major granule-binding proteins were found to present a pattern in two-dimensional electrophoresis that was very similar to but shifted slightly toward the basic end of the gel from the pattern generated by light chains associated with clathrin in adrenal medullary coated vesicles. In the chromaffin cell, these proteins, by associating with the granule membrane in the presence of an increased cytosolic Ca2+ concentration, might play a variety of roles in the process of exocytosis.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017310:Annexin A7; D002118:Calcium / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002837:Chromaffin Granules / Q000378:metabolism*; D002838:Chromaffin System / Q000378:metabolism*; D002846:Chromatography, Affinity; D007425:Intracellular Membranes / Q000378:metabolism; D050356:Lipid Metabolism; D008081:Liposomes / Q000378:metabolism; D008099:Liver / Q000032:analysis; D008854:Microscopy, Electron; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism
D017310:Annexin A7; D002352:Carrier Proteins; D008081:Liposomes; D011506:Proteins; D000255:Adenosine Triphosphate; D002118:Calcium