Regulation of alpha 1-acid glycoprotein externalization and intracellular accumulation in glucocorticoid-induced rat hepatoma cells.
alpha 1-Acid glycoprotein (alpha 1-AGP) is a glucocorticoid inducible gene product that is synthesized and secreted by certain rat hepatoma tissue culture (HTC) cell lines such as M1.54. Exposure to monensin, a Na+-K+ ionophore, causes a significant redistribution of alpha 1-AGP into two distinct fractions; immunoprecipitation of [35S]methionine-labeled proteins revealed that a 27% decrease in secretion accounts for a sixfold increase in accumulation of a stable intracellular species. The new intracellular alpha 1-AGP is more heterogeneous than normal while the extra-cellular form is 6000 Da smaller than normal. These effects are due to selective alterations in carbohydrate maturation; endo-beta-N-acetylglucosaminidase H (endo H) digestion demonstrated that both alpha 1-AGP species contain variable numbers of endo H-resistant oligosaccharide side chains ranging between zero and five. Ricin affinity chromatography revealed that the attachment of galactose residues is strikingly correlated with alpha 1-AGP externalization while neuraminidase digestions demonstrated that sialic acid attachment appears unessential for its secretion. Taken together, our results suggest that in the presence of monensin the cellular transport of intracellular destined and externalized alpha 1-AGP proceeds in common through the early segments of the Golgi and at a point prior to or at the compartment containing galactosyl transferase, alpha 1-AGP becomes committed for secretion.
Haffar
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Edwards
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Firestone
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D000118:Acetylglucosaminidase; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002846:Chromatography, Affinity; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008985:Monensin / Q000494:pharmacology; D009439:Neuraminidase; D009844:Oligosaccharides / Q000378:metabolism; D009961:Orosomucoid / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats; D012995:Solubility
D003902:Detergents; D009844:Oligosaccharides; D009961:Orosomucoid; D008985:Monensin; D009439:Neuraminidase; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase