Effect of inhibiting N-glycosylation on the stability and binding activity of the low density lipoprotein receptor.
Tunicamycin, a specific inhibitor of N-glycosylation, was used to study the function of asparagine-linked oligosaccharides of the low density lipoprotein (LDL) receptor in cultured human skin fibroblasts. When cells were preincubated in the presence of 0.5 micrograms/ml of the drug the incorporation of [3H]mannose into the receptor was completely prevented and that of [3H]glucosamine was reduced to approximately 41% of the control value. The [35S]methionine radioactivity detected in receptor core protein of tunicamycin-treated cells was about 52% of that measured in the receptor of control cells. The decrease in the radioactivity was similar in both the mature receptor as well as in its precursor form, and it was significantly greater than that found in total protein. The rates of receptor degradation in control- and tunicamycin-treated cells were comparable. Neither cell surface appearance of the newly synthesized LDL receptor nor its recycling were affected by tunicamycin. However, the LDL receptor produced in tunicamycin-treated cells was smaller in molecular size, and it exhibited an about 50% lower binding capacity when compared with its counterpart synthesized in control cells. This indicates that there is a relationship between N-glycosylation and the ligand binding activity of the LDL receptor. The possible role of asparagine-linked oligosaccharides in optimizing the biological activity of the LDL receptor is discussed.
Filipovic
I
I
Institute of Physiological Chemistry, University of Münster, Federal Republic of Germany.
D002478:Cells, Cultured; D004355:Drug Stability; D005347:Fibroblasts / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism; D011973:Receptors, LDL / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism*; D012867:Skin / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology
D008077:Lipoproteins, LDL; D011973:Receptors, LDL; D014415:Tunicamycin; D005944:Glucosamine; D008358:Mannose