Inhibition by 2-deoxy-D-glucose of synthesis of glycoprotein enzymes by protoplasts of Saccharomyces: relation to inhibition of sugar uptake and metabolism.
The synthesis of the glycoprotein enzymes, invertase and acid phosphatase, by protoplasts of Saccharomyces mutant 1016, is inhibited by 2-deoxy-d-glucose (2-dG) after a 20- to 30-min lag period under conditions (external sugar to 2-dG ratio of 40:1) which cause only a slight decrease in total protein synthesis. Formation of one intracellular enzyme, alpha-glucosidase, is also sensitive, but production of another, alkaline phosphatase, is unaffected. A nonmetabolized glucose analogue, 6-deoxy-d-glucose, had no inhibitory effect. The total uptake of external fructose and maltose was decreased by 2-dG after a lag period of about the same duration as that before the inhibition of synthesis of enzymes or of mannan and glucan; during this time 2-dG was taken up by the protoplasts and accumulated primarily as 2-dG-6-phosphate (2-dG-6-P). Studies in vitro showed that 2-dG-6-P inhibits both yeast phosphoglucose isomerase and phosphomannose isomerase. The intracellular levels of the 6-phosphates of glucose, fructose, and mannose did not increase in the presence of 2-dG. We suggest that the high internal level of 2-dG-6-P blocks synthesis of the cell wall polysaccharides and glycoproteins in two ways. It directly inhibits the conversion of fructose-6-P to glucose-6-P and to mannose-6-P. At the same time, it restricts the transport of fructose and maltose into the cell; however, the continuing limited uptake of the sugars still provides sufficient energy for protein synthesis. The cessation of alpha-glucosidase synthesis is probably a result of depletion of the internal pool of maltose (the inducer). Our findings support the suggestion that restriction of synthesis of the carbohydrate moiety of glycoproteins reduces formation of the active enzyme.
D000135:Acid Phosphatase / Q000096:biosynthesis*; D000469:Alkaline Phosphatase / Q000096:biosynthesis*; D050260:Carbohydrate Metabolism; D002247:Carbon Isotopes; D002473:Cell Wall / Q000378:metabolism; D003470:Culture Media; D005632:Fructose / Q000378:metabolism; D005958:Glucosephosphates; D005959:Glucosidases / Q000096:biosynthesis*; D006023:Glycoproteins / Q000096:biosynthesis; D006600:Hexosephosphates / Q000378:metabolism; D006601:Hexoses / Q000494:pharmacology*; D007535:Isomerases / Q000096:biosynthesis; D008320:Maltose / Q000378:metabolism; D010710:Phosphates / Q000494:pharmacology; D011134:Polysaccharides / Q000096:biosynthesis; D014176:Protein Biosynthesis; D011523:Protoplasts / Q000201:enzymology* / Q000378:metabolism; D012440:Saccharomyces / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D013393:Sucrase / Q000096:biosynthesis*; D013912:Threonine / Q000378:metabolism; D013997:Time Factors; D014316:Tritium
D002247:Carbon Isotopes; D003470:Culture Media; D005958:Glucosephosphates; D006023:Glycoproteins; D006600:Hexosephosphates; D006601:Hexoses; D010710:Phosphates; D011134:Polysaccharides; D014316:Tritium; D013912:Threonine; D005632:Fructose; D008320:Maltose; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D005959:Glucosidases; D013393:Sucrase; D007535:Isomerases