Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1972 Aug;111(2):419–429. doi: 10.1128/jb.111.2.419-429.1972

Inhibition by 2-Deoxy-d-Glucose of Synthesis of Glycoprotein Enzymes by Protoplasts of Saccharomyces: Relation to Inhibition of Sugar Uptake and Metabolism1

S-C Kuo a, J O Lampen a
PMCID: PMC251299  PMID: 5053466

Abstract

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.

Full text

PDF
419

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson J. S., Meadow P. M., Haskin M. A., Strominger J. L. Biosynthesis of the peptidoglycan of bacterial cell walls. I. Utilization of uridine diphosphate acetylmuramyl pentapeptide and uridine diphosphate acetylglucosamine for peptidoglycan synthesis by particulate enzymes from Staphylococcus aureus and Micrococcus lysodeikticus. Arch Biochem Biophys. 1966 Sep 26;116(1):487–515. doi: 10.1016/0003-9861(66)90056-7. [DOI] [PubMed] [Google Scholar]
  2. Azam F., Kotyk A. Glucose-6-phosphate as regulator of monosaccharide transport in baker's yeast. FEBS Lett. 1969 Mar;2(5):333–335. doi: 10.1016/0014-5793(69)80057-8. [DOI] [PubMed] [Google Scholar]
  3. Azam F., Kuo S. C., Cirillo V. P. Production of phenotypically epimeraseless yeast by L-arabinose. J Bacteriol. 1971 Jun;106(3):915–919. doi: 10.1128/jb.106.3.915-919.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biely P., Bauer S. The formation of guanosine diphosphate-2-deoxy-D-glucose in yeast. Biochim Biophys Acta. 1968 Mar 11;156(2):432–434. doi: 10.1016/0304-4165(68)90281-x. [DOI] [PubMed] [Google Scholar]
  5. Biely P., Krátký Z., Kovarík J., Bauer S. Effect of 2-deoxyglucose on cell wall formation in Saccharomyces cerevisiae and its relation to cell growth inhibition. J Bacteriol. 1971 Jul;107(1):121–129. doi: 10.1128/jb.107.1.121-129.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boer P., Steyn-Parvé E. P. Isolation and purification of an acid phosphatase from baker's yeast (Saccharomyces cerevisiae). Biochim Biophys Acta. 1966 Nov 15;128(2):400–402. doi: 10.1016/0926-6593(66)90189-5. [DOI] [PubMed] [Google Scholar]
  7. CIRILLO V. P. Mechanism of glucose transport across the yeast cell membrane. J Bacteriol. 1962 Sep;84:485–491. doi: 10.1128/jb.84.3.485-491.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cirillo V. P. Relationship between sugar structure and competition for the sugar transport system in Bakers' yeast. J Bacteriol. 1968 Feb;95(2):603–611. doi: 10.1128/jb.95.2.603-611.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DE LA FUENTE G., SOLS A. Transport of sugars in yeasts. II. Mechanisms of utilization of disaccharides and related glycosides. Biochim Biophys Acta. 1962 Jan 1;56:49–62. doi: 10.1016/0006-3002(62)90526-7. [DOI] [PubMed] [Google Scholar]
  10. Elorza M. V., Sentandreu R. Effect of cycloheximide on yeast cell wall synthesis. Biochem Biophys Res Commun. 1969 Aug 22;36(5):741–747. doi: 10.1016/0006-291x(69)90672-x. [DOI] [PubMed] [Google Scholar]
  11. Farkas V., Bauer S., Zemek J. Metabolis of 2-deoxyD-glucose in baker's yeast. 3. Formation of 2,2'-dideoxy-alpha, alpha'-trehalose. Biochim Biophys Acta. 1969 Jun 17;184(1):77–82. doi: 10.1016/0304-4165(69)90100-7. [DOI] [PubMed] [Google Scholar]
  12. Farkas V., Svoboda A., Bauer S. Inhibitory effect of 2-deoxy-d-glucose on the formation of the cell wall in yeast protoplasts. J Bacteriol. 1969 May;98(2):744–748. doi: 10.1128/jb.98.2.744-748.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farkas V., Svoboda A., Bauer S. Secretion of cell-wall glycoproteins by yeast protoplasts. Effect of 2-deoxy-D-glucose and cycloheximide. Biochem J. 1970 Aug;118(5):755–758. doi: 10.1042/bj1180755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Görts C. P. Effect of glucose on the activity and the kinetics of the maltose-uptake system and of alpha-glucosidase in Saccharomyces cerevisiae. Biochim Biophys Acta. 1969 Jul 30;184(2):299–305. doi: 10.1016/0304-4165(69)90032-4. [DOI] [PubMed] [Google Scholar]
  15. HALVORSON H., ELLIAS L. The purification and properties of an alpha-glucosidase of Saccharomyces italicus Y1225. Biochim Biophys Acta. 1958 Oct;30(1):28–40. doi: 10.1016/0006-3002(58)90237-3. [DOI] [PubMed] [Google Scholar]
  16. HEREDIA C. F., DELAFUENTE G., SOLS A. METABOLIC STUDIES WITH 2-DEOXYHEXOSES. I. MECHANISMS OF INHIBITION OF GROWTH AND FERMENTATION IN BAKER'S YEAST. Biochim Biophys Acta. 1964 May 11;86:216–223. doi: 10.1016/0304-4165(64)90045-5. [DOI] [PubMed] [Google Scholar]
  17. HEREDIA C. F., YEN F., SOLS A. Role and formation of the acid phosphatase in yeast. Biochem Biophys Res Commun. 1963 Jan 18;10:14–18. doi: 10.1016/0006-291x(63)90259-6. [DOI] [PubMed] [Google Scholar]
  18. Heredia C. F., Sols A., DelaFuente G. Specificity of the constitutive hexose transport in yeast. Eur J Biochem. 1968 Aug;5(3):321–329. doi: 10.1111/j.1432-1033.1968.tb00373.x. [DOI] [PubMed] [Google Scholar]
  19. Johnson B. F. Lysis of yeast cell walls induced by 2-deoxyglucose at their sites of glucan synthesis. J Bacteriol. 1968 Mar;95(3):1169–1172. doi: 10.1128/jb.95.3.1169-1172.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. KIPNIS D. M., CORI C. F. Studies of tissue permeability. V. The penetration and phosphorylation of 2-deoxyglucose in the rat diaphragm. J Biol Chem. 1959 Jan;234(1):171–177. [PubMed] [Google Scholar]
  21. Kotyk A. Properties of the sugar carrier in baker's yeast. II. Specificity of transport. Folia Microbiol (Praha) 1967;12(2):121–131. doi: 10.1007/BF02896872. [DOI] [PubMed] [Google Scholar]
  22. Kuo S. C., Lampen J. O. Osmotic regulation of invertase formation and secretion by protoplasts of Saccharomyces. J Bacteriol. 1971 Apr;106(1):183–191. doi: 10.1128/jb.106.1.183-191.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lampen J. O. External enzymes of yeast: their nature and formation. Antonie Van Leeuwenhoek. 1968;34(1):1–18. doi: 10.1007/BF02046409. [DOI] [PubMed] [Google Scholar]
  24. Liras P., Gascón S. Biosynthesis and secretion of yeast invertase. Effect of cycloheximide and 2-deoxy-D-glucose. Eur J Biochem. 1971 Nov 11;23(1):160–165. doi: 10.1111/j.1432-1033.1971.tb01603.x. [DOI] [PubMed] [Google Scholar]
  25. MCLELLAN W. L., Jr, LAMPEN J. O. The acid phosphatase of yeast. Localization and secretion by protoplasts. Biochim Biophys Acta. 1963 Feb 12;67:324–326. doi: 10.1016/0006-3002(63)91832-8. [DOI] [PubMed] [Google Scholar]
  26. NORTHCOTE D. H., HORNE R. W. The chemical composition and structure of the yeast cell wall. Biochem J. 1952 May;51(2):232–236. doi: 10.1042/bj0510232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neumann N. P., Lampen J. O. Purification and properties of yeast invertase. Biochemistry. 1967 Feb;6(2):468–475. doi: 10.1021/bi00854a015. [DOI] [PubMed] [Google Scholar]
  28. PALADINI A. C., LELOIR L. F. Studies on uridine-diphosphate-glucose. Biochem J. 1952 Jun;51(3):426–430. doi: 10.1042/bj0510426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. ROBERTSON J. J., HALVORSON H. O. The components of maltozymase in yeast, and their behavior during deadaptation. J Bacteriol. 1957 Feb;73(2):186–198. doi: 10.1128/jb.73.2.186-198.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. SCHMIDT G., BARTSCH G., LAUMONT M. C., HERMAN T., LISS M. Acid phosphatase of bakers' yeast: an enzyme of the external cell surface. Biochemistry. 1963 Jan-Feb;2:126–131. doi: 10.1021/bi00901a022. [DOI] [PubMed] [Google Scholar]
  31. SUTTON D. D., LAMPEN J. O. Localization of sucrose and maltose fermenting systems in Saccharomyces cerevisiae. Biochim Biophys Acta. 1962 Jan 29;56:303–312. doi: 10.1016/0006-3002(62)90567-x. [DOI] [PubMed] [Google Scholar]
  32. Steiner S., Lester R. L. Studies on the diversity of inositol-containing yeast phospholipids: incorporation of 2-deoxyglucose into lipid. J Bacteriol. 1972 Jan;109(1):81–88. doi: 10.1128/jb.109.1.81-88.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. TORRIANI A. Influence of inorganic phosphate in the formation of phosphatases by Escherichia coli. Biochim Biophys Acta. 1960 Mar 11;38:460–469. doi: 10.1016/0006-3002(60)91281-6. [DOI] [PubMed] [Google Scholar]
  34. Van Wijk R., Ouwehand J., van den Bos T., Koningsberger V. V. Induction and catabolite repression of alpha-glucosidase synthesis in protoplasts of Saccharomyces carlsbergensis. Biochim Biophys Acta. 1969 Jul 22;186(1):178–191. doi: 10.1016/0005-2787(69)90501-2. [DOI] [PubMed] [Google Scholar]
  35. WEIMBERG R., ORTON W. L. REPRESSIBLE ACID PHOSPHOMONOESTERASE AND CONSTITUTIVE PYROPHOSPHATASE OF SACCHAROMYCES MELLIS. J Bacteriol. 1963 Oct;86:805–813. doi: 10.1128/jb.86.4.805-813.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. WICK A. N., DRURY D. R., NAKADA H. I., WOLFE J. B. Localization of the primary metabolic block produced by 2-deoxyglucose. J Biol Chem. 1957 Feb;224(2):963–969. [PubMed] [Google Scholar]
  37. van Wijk R. Alpha-glucosidase synthesis, respiratory enzymes and catabolite repression in yeast. I. The effects of glucose and maltose on inducible alpha-glucosidase synthesis in protoplasts of S. carlsbergensis. Proc K Ned Akad Wet C. 1968;71(1):60–71. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES