Differential subcellular distribution of p36 (the heavy chain of calpactin I) and other annexins in the adrenal medulla.
The annexins are a group of highly related Ca2(+)-dependent membrane-binding proteins that are present in a wide variety of cells and tissues. We have examined the subcellular distribution of five members of the annexin family in the adrenal medulla. Bovine adrenal medullary tissue was homogenized in buffers containing EGTA and fractionated on sucrose gradients. p36 (the large subunit of calpactin I) was found to be predominantly membrane associated, with approximately 20% present in fractions enriched in chromaffin granules. In contrast, lipocortin I was localized primarily to the cytosol, with only a small proportion found in plasma membrane-containing fractions. Like lipocortin I, endonexin I was found to be present almost entirely in the soluble fractions. The 67-kDa calelectrin was localized primarily to the plasma membrane fractions, with a small amount present in the chromaffin granule and cytoplasmic fractions. Synexin was present in both membranous and cytoplasmic fractions. p36 appeared to be a peripherally associated granule membrane protein in that it was dissociated from the membrane by addition of base and it partitioned with the aqueous phase when granule membranes were treated with Triton X-114. Antiserum against p10 (the small subunit of calpactin I) reacted with a protein of 19 kDa that is specifically localized in chromaffin granule membrane fractions. The differences in subcellular distributions of the annexins suggest that these proteins have distinct cellular functions. The finding that p36 is associated with chromaffin granule and plasma membrane fractions provides further support for a possible role of calpactin in exocytosis.
Drust
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DS
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville 22908.
Creutz
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CE
D000313:Adrenal Medulla / Q000737:chemistry / Q000648:ultrastructure*; D000818:Animals; D017304:Annexin A5; D017310:Annexin A7; D017302:Annexins; D002135:Calcium-Binding Proteins / Q000032:analysis*; D002417:Cattle; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry; D002499:Centrifugation, Density Gradient; D002837:Chromaffin Granules / Q000737:chemistry; D003600:Cytosol / Q000737:chemistry; D007425:Intracellular Membranes / Q000737:chemistry; D011506:Proteins / Q000032:analysis; D013347:Subcellular Fractions / Q000737:chemistry*
D017304:Annexin A5; D017310:Annexin A7; D017302:Annexins; D002135:Calcium-Binding Proteins; D011506:Proteins