Endocrine secretory granules and neuronal synaptic vesicles have three integral membrane proteins in common.
In response to an external stimulus, neuronal cells release neurotransmitters from small synaptic vesicles and endocrine cells release secretory proteins from large dense core granules. Despite these differences, endocrine cells express three proteins known to be components of synaptic vesicle membranes. To determine if all three proteins, p38, p65, and SV2, are present in endocrine dense core granule membranes, monoclonal antibodies bound to beads were used to immunoisolate organelles containing the synaptic vesicle antigens. [3H]norepinephrine was used to label both chromaffin granules purified from the bovine adrenal medulla and rat pheochromocytoma (PC12) cells. Up to 80% of the vesicular [3H]norepinephrine was immunoisolated from both labeled purified bovine chromaffin granules and PC12 postnuclear supernatants. In PC12 cells transfected with DNA encoding human growth hormone, the hormone was packaged and released with norepinephrine. 90% of the sedimentable hormone was also immunoisolated by antibodies to all three proteins. Stimulated secretion of PC12 cells via depolarization with 50 mM KCl decreased the amount of [3H]norepinephrine or human growth hormone immunoisolated. Electron microscopy of the immunoisolated fractions revealed large (greater than 100 nm diameter) dense core vesicles adherent to the beads. Thus, large dense core vesicles containing secretory proteins possess all three of the known synaptic vesicle membrane proteins.
Lowe
A W
AW
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448.
Madeddu
L
L
Kelly
R B
RB
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002837:Chromaffin Granules / Q000032:analysis; D003594:Cytoplasmic Granules / Q000032:analysis*; D007158:Immunologic Techniques; D007425:Intracellular Membranes / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis*; D008854:Microscopy, Electron; D009638:Norepinephrine / Q000378:metabolism; D051381:Rats; D013572:Synaptic Vesicles / Q000032:analysis*
D008565:Membrane Proteins; D009638:Norepinephrine