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. 1971 Apr;106(1):183–191. doi: 10.1128/jb.106.1.183-191.1971

Osmotic Regulation of Invertase Formation and Secretion by Protoplasts of Saccharomyces

S-C Kuo 1, J O Lampen 1
PMCID: PMC248660  PMID: 5551635

Abstract

Osmotic regulation of invertase formation and secretion by protoplasts of Saccharomyces was examined using sorbitol, KCl, NaCl, or magnesium sulfate as the osmotic support. The synthesis and secretion of the enzyme is remarkably sensitive to the osmolarity of the supporting medium irrespective of the particular support employed. Invertase formation was inhibited at high osmolarity and was maximal at 0.65 to 0.75 osmolal, even though some leakage of the intracellular enzyme α-glucosidase and of ultraviolet (UV)-absorbing materials occurred under these conditions. The reduction of invertase formation and secretion due to high osmolarity was eliminated promptly when protoplasts were transferred into a medium of lower osmolarity. The rate of fructose uptake and of threonine incorporation into protein was decreased by high osmolarity; also reduction of invertase formation could be partially reversed by increasing the level of sugar supplied as energy source. Thus changes in the permeability of the plasma membrane (and presumably also in its structure) are important factors in the response of protoplasts to high osmolarity, though certainly not the complete explanation. Protoplasts suspended in 0.8 m sorbitol, with 10mm fructose as the energy source, increased their invertase level 5- to 10-fold during a 2-hr incubation without substantial release of α-glucosidase or UV-absorbing materials. Both the large and small forms of invertase were present in the protoplasts, but only the large form was released into the medium when enzyme was being actively synthesized. Formation and secretion of newly formed invertase and the release of enzyme initially present were inhibited by cycloheximide.

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Selected References

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  1. DAVIES A. Invertase formation in Saccharomyces fragilis. J Gen Microbiol. 1956 Feb;14(1):109–121. doi: 10.1099/00221287-14-1-109. [DOI] [PubMed] [Google Scholar]
  2. DODYK F., ROTHSTEIN A. FACTORS INFLUENCING THE APPEARANCE OF INVERTASE IN SACCHAROMYCES CEREVISIAE. Arch Biochem Biophys. 1964 Mar;104:478–486. doi: 10.1016/0003-9861(64)90492-8. [DOI] [PubMed] [Google Scholar]
  3. Gascón S., Lampen J. O. Purification of the internal invertase of yeast. J Biol Chem. 1968 Apr 10;243(7):1567–1572. [PubMed] [Google Scholar]
  4. Gascón S., Ottolenghi P. Invertase isozymes and their localization in yeast. C R Trav Lab Carlsberg. 1967;36(5):85–93. [PubMed] [Google Scholar]
  5. 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]
  6. Hutchison H. T., Hartwell L. H. Macromolecule synthesis in yeast spheroplasts. J Bacteriol. 1967 Nov;94(5):1697–1705. doi: 10.1128/jb.94.5.1697-1705.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. MILLBANK J. W. DNA AND PROTEIN SYNTHESIS IN THE GROWING YEAST PROTOPLAST. Exp Cell Res. 1964 Jun;35:77–83. doi: 10.1016/0014-4827(64)90073-4. [DOI] [PubMed] [Google Scholar]
  10. McKeehan W., Hardesty B. The mechanism of cycloheximide inhibition of protein synthesis in rabbit reticulocytes. Biochem Biophys Res Commun. 1969 Aug 15;36(4):625–630. doi: 10.1016/0006-291x(69)90351-9. [DOI] [PubMed] [Google Scholar]
  11. McMurrough I., Rose A. H. Effect of growth rate and substrate limitation on the composition and structure of the cell wall of Saccharomyces cerevisiae. Biochem J. 1967 Oct;105(1):189–203. doi: 10.1042/bj1050189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. SIEGEL M. R., SISLER H. D. SITE OF ACTION OF CYCLOHEXIMIDE IN CELLS OF SACCHAROMYCES PASTORIANUS. II. THE NATURE OF INHIBITION OF PROTEIN SYNTHESIS IN A CELL-FREE SYSTEM. Biochim Biophys Acta. 1964 May 18;87:83–89. [PubMed] [Google Scholar]
  14. SMITH L. Structure of the bacterial respiratory-chain system. Respiration of Bacillus subtilis spheroplasts as a function of the osmotic pressure of the medium. Biochim Biophys Acta. 1962 Jul 30;62:145–152. doi: 10.1016/0006-3002(62)90499-7. [DOI] [PubMed] [Google Scholar]
  15. Trevithick J. R., Metznberg R. L. The invertase isozyme formed by Neurospora protoplasts. Biochem Biophys Res Commun. 1964 Jul 1;16(4):319–325. doi: 10.1016/0006-291x(64)90033-6. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. de KLOET S., van WERMESKERKEN R., KONINGSBERGER V. V. Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis. I. The effect of ribonuclease on protein synthesis. Biochim Biophys Acta. 1961 Feb 12;47:138–143. doi: 10.1016/0006-3002(61)90838-1. [DOI] [PubMed] [Google Scholar]
  18. de Kloet S. R. Accumulation of RNA with a DNA like base composition in Saccharomyces carlsbergensis in the presence of cycloheximide. Biochem Biophys Res Commun. 1965 May 18;19(5):582–586. doi: 10.1016/0006-291x(65)90378-5. [DOI] [PubMed] [Google Scholar]
  19. de Kloet S. R. Ribonucleic acid synthesis in yeast. The effect of cycloheximide on the synthesis of ribonucleic acid in Saccharomyces carlsbergensis. Biochem J. 1966 Jun;99(3):566–581. doi: 10.1042/bj0990566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]

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