Targeting of the zymogen-granule protein syncollin in AR42J and AtT-20 cells.
Syncollin is a 13-kDa protein associated with the membranes of pancreatic zymogen granules. Here we determine the in situ localization of syncollin in pancreatic acinar cells from adult and neonatal rats, and study the targeting of green fluorescent protein-(GFP-) and His(6)-tagged syncollin chimaeras in model exocrine and endocrine secretory cells. Immunocytochemical analysis of the distribution of syncollin in fully differentiated and neonatal acinar cells revealed a granular pattern that corresponded with that of the zymogen-granule markers synaptobrevin 2 and amylase. In fully differentiated acinar cells syncollin-positive vesicles were detected in the apical region of the cells, whereas in neonatal acinar cells they were found clustered near the cell nucleus. Both GFP- and His(6)-tagged syncollin entered the secretory pathway when transiently expressed in AR42J or AtT-20 cells. Syncollin-GFP was found predominantly in amylase-positive granules in AR42J cells and in adrenocorticotrophic hormone- (ACTH-) positive granules in AtT-20 cells. Syncollin-GFP was also present in the Golgi complex in AR42J cells. Syncollin-His(6) became localized in ACTH-containing granules in the neuritic processes of AtT-20 cells. In AR42J cells syncollin-His(6) did not co-localize with amylase, but was detected in acidic vesicles. These results show that the exocrine protein syncollin contains intrinsic cell-type-independent targeting information that is retained in both exocrine and endocrine cells after fusion to the GFP tag. In contrast, His(6)-tagged syncollin is efficiently targeted to secretory granules only in AtT-20 cells and not in AR42J cells.
Hodel
A
A
Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QJ, U.K.
Edwardson
J M
JM
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D004792:Enzyme Precursors / Q000378:metabolism*; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D010179:Pancreas / Q000166:cytology / Q000378:metabolism; D051381:Rats
D002352:Carrier Proteins; D004792:Enzyme Precursors; D008565:Membrane Proteins; C107454:Sycn protein, rat