Inhibition of glucose trimming by castanospermine results in rapid degradation of unassembled major histocompatibility complex class I molecules.
The CMT-cKd1 cell line provides a system for studying the initial processing steps of N-linked oligosaccharides as these cells have been shown to produce major histocompatibility complex (MHC) class I molecules which, due to a defect in assembly, recycle between the endoplasmic reticulum and a pre-Golgi compartment, failing to reach the cell surface (Hsu, V.W., Yuan, L. C., Nuchtern, J. G., Lippincott-Schwartz, J., Hämmerling, G. J., and Klausner, R. D. (1991) Nature 352, 441-444). In the present study we observed that when the MHC class I heavy chain of these CMT cells was pulse-radiolabeled with [35S]methionine in the presence of the glucosidase inhibitor, castanospermine (CST), it underwent a rapid degradation during a 60-min chase, in contrast to control cells in which it remained stable during that period. The CST-promoted instability of the MHC molecule appeared to be specific, as it did not occur when 1-deoxymannojirimycin, an inhibitor of mannosidase, was added to the cells. Although endomannosidase was found to be present in the CMT cells, the electrophoretic mobility of the MHC heavy chain produced in the presence of CST indicated that deglucosylation through the alternate route provided by this enzyme did not occur. Furthermore, gamma-interferon did not prevent the rapid disappearance of the MHC molecule, although it brought about entry of this glycoprotein into the secretory pathway in cells incubated without CST. The results of our studies suggest that retention of glucose on N-linked oligosaccharides may under certain circumstances provide a signal for pre-Golgi protein degradation.
Moore
S E
SE
Department of Biological Chemistry, Harvard Medical School, Boston, Massachusetts.
Spiro
R G
RG
D017485:1-Deoxynojirimycin / Q000494:pharmacology; D002478:Cells, Cultured; D005947:Glucose / Q000378:metabolism*; D005959:Glucosidases / Q000037:antagonists & inhibitors*; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D007212:Indolizines / Q000494:pharmacology*; D054836:Indolizidines
D017485:1-Deoxynojirimycin; D005947:Glucose; D005959:Glucosidases; D015395:Histocompatibility Antigens Class I; D007212:Indolizines; C037806:castanospermine; D054836:Indolizidines