Chromogranin B (secretogranin I), a secretory protein of the regulated pathway, is also present in a tightly membrane-associated form in PC12 cells.
Chromogranin B (CgB, also called secretogranin I) is a secretory protein sorted to secretory granules in a wide variety of endocrine cells and neurons. Unexpectedly, after stimulation of regulated secretion in the neuroendocrine cell line PC12, a fraction of the exocytosed CgB was not released into the medium but remained associated with the plasma membrane. The addition of exogenous CgB to unstimulated cells did not result in the appearance of cell surface CgB, suggesting that the presence of cell surface CgB could not be accounted for by adsorption of released CgB to the cell surface. Upon further incubation of stimulated PC12 cells, the surface CgB was internalized by the cells and largely degraded. The surface CgB was not released by exposure to pH 11, yet it partitioned in the aqueous phase upon Triton X-114 phase separation. Subcellular fractionation and differential extraction studies showed that the membrane-associated CgB constituted at least 10% of the total cellular CgB. These observations suggest that (a) the appearance of CgB at the cell surface is due to fusion of secretory granules with the plasma membrane and (b) a fraction of CgB is present in tight association with the secretory granule membrane. We propose a model in which membrane-associated CgB, by virtue of its ability to interact in a homophilic manner with soluble CgB, plays a key role in the sorting and targeting of CgB to the regulated pathway.
Pimplikar
S W
SW
Cell Biology Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
Huttner
W B
WB
D000818:Animals; D001710:Biotin / Q000378:metabolism; D015153:Blotting, Western; D002458:Cell Fractionation; D002864:Chromogranins / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D005455:Fluorescent Antibody Technique; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D016716:PC12 Cells; D011233:Precipitin Tests
D002864:Chromogranins; D008565:Membrane Proteins; D001710:Biotin