Glucose starvation leads in rat hepatoma cells to partially N-glycosylated glycoproteins including alpha 1-acid glycoproteins. Identification by endoglycolytic digestions in polyacrylamide gels.
Within minutes of glucose starvation confluent monolayers of rat hepatoma cells synthesize glycoproteins, including alpha 1-acid glycoprotein, which appear on two-dimensional gels as size heterogeneous spot series. The longer the period of glucose starvation the more the production of the glycoproteins is shifted toward smaller molecular weight forms. To compare these forms with the corresponding glycoproteins synthesized either in a cell-free system or by nonstarved cells, a mapping of the N-glycan was done by endo-beta-N-acetylglucosaminidase digestion within a polyacrylamide gel. Glycoproteins from glucose-starved cells contain a reduced number of N-glycans which belong to both the endo H-sensitive and resistant type. The decrease of N-glycosylation may be correlated with the accumulation of truncated lipid-bound oligosaccharides, for the gel chromatography of the oligosaccharides released from the lipid and protein fractions of glucose-starved cells revealed a drastic reduction in their size. Most of the lipid-linked oligosaccharides synthesized during glucose starvation are resistant to endo H digestion. Under conditions of limiting glycosylation we were able to show by glycopeptide analysis, that in the case of alpha 1-acid glycoprotein, N-glycans are added randomly to the 6 possible N-glycosylation sites. Furthermore, non- or partially N-glycosylated proteins do not acquire additional oligosaccharide units after restoration of glucose although the proteins can undergo secondary modification and, in the case of the secretory proteins, can be exported.
D000118:Acetylglucosaminidase; D000818:Animals; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005947:Glucose / Q000378:metabolism*; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D006027:Glycosides / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D009961:Orosomucoid / Q000378:metabolism*; D051381:Rats
D006023:Glycoproteins; D006027:Glycosides; D009961:Orosomucoid; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D005947:Glucose