Cathepsin D and beta-hexosaminidase synthesized in the presence of 1-deoxynojirimycin accumulate in the endoplasmic reticulum.
Biosynthesis, transport, and maturation of cathepsin D and beta-hexosaminidase was examined in fibroblasts exposed to 1-deoxynojirimycin, a glucose analogue known to inhibit trimming glucosidases (Saunier, B., Kilker, R. D., Jr., Tkacz, J. S., Quaroni, A., and Herscovics, A. (1982) J. Biol. Chem. 257, 14155-14161; Hettkamp, H., Bause, E., and Legler, G. (1982) Biosci. Rep. 2, 899-906). Cells treated with 1-deoxynojirimycin contained precursors of cathepsin D and beta-hexosaminidase larger by about 1-2 kDa than control cells. The shift in molecular size was probably due to glucose residues that were rapidly removed from the precursors in the absence but not in the presence of 1-deoxynojirimycin. In addition, 1-deoxynojirimycin inhibited the glycosylation of the beta-chain precursor of beta-hexosaminidase and the synthesis of glycoproteins, including that of cathepsin D. The proteolytic processing of the larger precursors was retarded by several hours. The delay in proteolytic maturation was secondary to the accumulation of the larger precursors in organelles, which fractionated with membranes of the endoplasmic reticulum and Golgi complex. The accumulated cathepsin D precursor contained neither mannose 6-phosphate residues nor complex type oligosaccharides, which are formed in the cis and trans aspects of the Golgi complex. Cathepsin D precursors eventually released from the site of accumulation were apparently deglucosylated, acquired mannose 6-phosphate residues and complex type oligosaccharides, and were transferred into lysosomes as efficiently as in control cells. Our results suggest that transport of cathepsin D from the endoplasmic reticulum to the Golgi complex depends on removal of glucose residues from its carbohydrate.
D017485:1-Deoxynojirimycin; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002402:Cathepsin D; D002403:Cathepsins / Q000096:biosynthesis* / Q000378:metabolism; D002458:Cell Fractionation; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D005347:Fibroblasts / Q000201:enzymology; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006596:Hexosaminidases / Q000096:biosynthesis* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D001619:beta-N-Acetylhexosaminidases
D000900:Anti-Bacterial Agents; D017485:1-Deoxynojirimycin; D006596:Hexosaminidases; D001619:beta-N-Acetylhexosaminidases; D002403:Cathepsins; D002402:Cathepsin D; D005944:Glucosamine