Polarized secretion of the regulated secretory protein chromogranin A.
Bovine chromogranin A (CgA), together with secreted alkaline phosphatase (SEAP) as an external control for apical secretion were expressed in MDCK cells to test if CgA contains sorting signals for polarized secretion. CgA, SEAP, and the endogenous apical marker GP80 were secreted 75-80% apically. Basolateral secretion of SEAP was inhibited 40% by ammonium chloride. Sulfate labeling and digestion with chondroitinase ABC revealed a 120 kDa proteoglycan-CgA and 75 kDa CgA. Inhibition of proteoglycan synthesis did not affect apical secretion of CgA. As CgA is not N-glycosylated, we used tunicamycin to test if cellular N-glycosylation is required for apical sorting. Tunicamycin reversed the polarity of secretion of CgA to the basolateral side. These results suggest that CgA contains dominant apical and recessive basolateral sorting information.
Kühn
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Department of Molecular, Cellular, and Craniofacial Biology, University of Louisville, Louisville, Kentucky 40292, USA.
Cohn
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Gorr
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D000818:Animals; D050260:Carbohydrate Metabolism; D002417:Cattle; D002460:Cell Line; D002809:Chondroitin Sulfates / Q000378:metabolism; D053379:Chromogranin A; D002864:Chromogranins / Q000378:metabolism*; D004285:Dogs; D004705:Endocytosis; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000166:cytology / Q000378:metabolism
D053379:Chromogranin A; D002864:Chromogranins; D002809:Chondroitin Sulfates