title: Reconstitution in vitro of the pH-dependent aggregation of pancreatic zymogens en route to the secretory granule: implication of GP-2.
abstract: Regulated secretory proteins are thought to be sorted in the trans-Golgi network (TGN) via selective aggregation. To elucidate the biogenesis of the secretory granule in the exocrine pancreas, we reconstituted in vitro the conditions of pH and ions believed to exist in the TGN using the end product of this sorting process, the zymogen granule contents. Protein aggregation was dependent on pH (acidic) and on the presence of cations (10 mM Ca2+, 150 mM K+) to reproduce the pattern of proteins found in the granule. The constitutive secretory protein IgG was excluded from these aggregates. Zymogen aggregation correlated with the relative proportion of the major granule membrane protein GP-2 in the assay. These results show that the glycosylphosphatidylinositol-anchored protein GP-2 co-aggregates with zymogens in the acidic environment believed to exist in the pancreatic TGN, and thus suggest that GP-2 would function as a membrane anchor for zymogen aggregates, facilitating their entrapment in budding vesicles directed towards the regulated secretory pathway.
authors: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Québec, Canada.
authors: F A
authors: FA
authors: Leblond
authors: G
authors: G
authors: Viau
authors: J
authors: J
authors: Lainé
authors: D
authors: D
authors: Lebel
mesh_terms: D002118:Calcium / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000201:enzymology*; D004792:Enzyme Precursors / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D009994:Osmolar Concentration; D010179:Pancreas / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure
chemical_list: D004792:Enzyme Precursors; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; C072961:glycoprotein-2, pancreas; D002118:Calcium