Syncollin inhibits regulated corticotropin secretion from AtT-20 cells through a reduction in the secretory vesicle population.
Syncollin is a 13 kDa protein that is highly expressed in the exocrine pancreas. Syncollin normally exists as a doughnut-shaped homo-oligomer (quite probably a hexamer) in close association with the luminal surface of the zymogen granule membrane. In the present study, we examine the effect of expression of syncollin in AtT-20 neuroendocrine cells, which do not normally express this protein. Efficient expression was achieved by infection of the cells with adenoviral constructs encoding either untagged or GFP (green fluorescent protein)-tagged syncollin. Both forms of the protein were sorted into corticotropin (ACTH)-positive secretory vesicles present mainly at the tips of cell processes. Neither form affected basal corticotropin secretion or the constitutive secretion of exogenously expressed secreted alkaline phosphatase. In contrast, regulated secretion of corticotropin was inhibited (by 49%) by untagged but not by GFP-tagged syncollin. In parallel, untagged syncollin caused a 46% reduction in the number of secretory vesicles present at the tips of the cell processes. Syncollin-GFP was without effect. We could also show that native syncollin purified from rat pancreas was capable of permeabilizing erythrocytes. We suggest that syncollin may induce uncontrolled permeabilization of corticotropin-containing vesicles and subsequently destabilize them. Both forms of syncollin were tightly membrane-associated and appeared to exist as homooligomers. Hence, the lack of effect of syncollin-GFP on regulated exocytosis suggests that the GFP tag interferes in a subtler manner with the properties of the assembled protein.
Wäsle
Barbara
B
Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK.
Hays
Lori B
LB
Rhodes
Christopher J
CJ
Edwardson
J Michael
JM
D000324:Adrenocorticotropic Hormone / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000502:physiology*; D002460:Cell Line; D006801:Humans; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D009474:Neurons / Q000378:metabolism; D009490:Neurosecretory Systems / Q000166:cytology / Q000378:metabolism*; D022142:Secretory Vesicles / Q000378:metabolism*
D002352:Carrier Proteins; D008565:Membrane Proteins; C107454:Sycn protein, rat; D000324:Adrenocorticotropic Hormone