Passive sorting in maturing granules of AtT-20 cells: the entry and exit of salivary amylase and proline-rich protein.
Previous studies have suggested that salivary amylase and proline-rich protein are sorted differently when expressed in AtT-20 cells (Castle, A.M., L.E. Stahl, and J.D. Castle. 1992. J. Biol. Chem. 267:13093- 13100; Colomer, V., K. Lal, T.C. Hoops, and M.J. Rindler. 1994.EMBO (Eur. Mol. Biol. Organ.) J. 13:3711- 3719). We now show that both exocrine proteins behave similarly and enter the regulated secretory pathway as judged by immunolocalization and secretagogue- dependent stimulation of secretion. Analysis of stimulated secretion of newly synthesized proline-rich protein, amylase, and endogenous hormones indicates that the exogenous proteins enter the granule pool with about the same efficiency as the endogenous hormones. However, in contrast to the endogenous hormones, proline-rich protein and amylase are progressively removed from the granule pool during the process of granule maturation such that only small portions remain in mature granules where they colocalize with the stored hormones. The exogenous proteins that are not stored are recovered from the incubation medium and are presumed to have undergone constitutive-like secretion. These results point to a level of sorting for regulated secretion after entry of proteins into forming granules and indicate that retention is essential for efficient storage. Consequently, the critical role of putative sorting receptors for regulated secretion may be in retention rather than in granule entry.
Castle
A M
AM
Department of Cell Biology, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA.
Huang
A Y
AY
Castle
J D
JD
D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D000681:Amylases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D003594:Cytoplasmic Granules / Q000378:metabolism*; D004558:Electric Stimulation; D005089:Exocytosis; D006056:Golgi Apparatus / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D051379:Mice; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D055232:Proline-Rich Protein Domains; D051381:Rats; D012471:Salivary Proteins and Peptides / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D010455:Peptides; D012471:Salivary Proteins and Peptides; D000324:Adrenocorticotropic Hormone; D000681:Amylases