Abstract
Intact vacuoles are released from spheroplasts of Saccharomyces cerevisiae by means of a gentle mechanical disintegration method. They are purified by centrifugation in isotonic density gradients (flotation and subsequent sedimentation), and analyzed for their soluble amino acid content. The results indicate that about 60% of the total amino acid pool of spheroplasts is contained in the vacuoles. This may be an underestimate, as it presupposes no loss of amino acids from the vacuoles during the purification procedure. The amino acid concentration in the vecuoles is calculated to be approximately 5 times that in the cytoplasm if the total volumes of the two compartments are used for the calculation. The vacuolar amino acid pool is rich in basic amino acids, and in citrulline and glutamine, but contains a remarkably small amount of glutamate. Radioactive labeling experiments with spheroplasts indicate that the vacuolar amino acids are separated from the metabolically active pools located in the cytoplasm. This is particularly evident for the basic amino acids and glutamine; in contrast, the neutral amino acids and glutamate appear to exchange more rapidly between the cytoplasmic and the vacuolar compartments of the cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Atkinson, D. E.: Limitation of metabolite concentration and the conservation of solvent capacity in the living cell. In: Current topics in cellular regulation, Vol. 1, B. L. Horecker and E. R. Stadtman, Eds., pp. 29–43. New York: Academic Press 1969
Bourgeois, C.: Influence de la lysine sur la croissance de Saccharomyces cerevisiae. Bull. Soc. Chim. biol. (Paris) 51, 935–949 (1969)
Brown, C. M., Stanley, S. O.: Environment-mediated changes in the cellular content of the “pool” constituents and their associated changes in cell physiology. J. appl. Chem. Biotechnol. 22, 363–389 (1972)
Cercek, L., Cercek, B.: Relationship between changes in the structuredness of cytoplasm and rate constants for the hydrolysis of FDA in Saccharomyces cerevisiae. Biophysik 9, 109–112 (1973)
Conway, E. J., Armstrong, W. D.: The total intracellular concentration of solutes in yeast and other plant cells and the distensibility of the plant cell wall. Biochem. J. 81, 631–639 (1961)
Crabeel, M., Grenson, M.: Regulation of histidine uptake by specific feedback inhibition of two histidine permeases in Saccharomyces cerevisiae. Europ. J. Biochem. 14, 197–204 (1970)
Croes, A. F.: Induction of meiosis in yeast: Timing of cytological and biochemical events. Planta (Berl.) 76, 209–226 (1967)
Damadian, R.: Biological ion exchange resins. Ann. N. Y. Acad. Sci. 204, 211–248 (1973)
Davis, R. H.: Metabolite distribution. Science 178, 840–885 (1972)
Ferguson, A. R., Sims, A. P.: The regulation of glutamine metabolism in Candida utilis: the role of glutamine in the control of glutamine synthetase. J. gen. Microbiol. 80, 159–171 (1974)
Gancedo, J. M., Gancedo, C.: Concentrations of intermediary metabolites in yeast. Biochemistry 55, 205–211 (1973)
Guilliermond, A., Gautheret, R.: Recherches sur la coloration vitale des cellules végétales. Rev. gen. Bot. 53, 275–289 (1946)
Holden, J. T.: The composition of microbial amino acid pools. In: Amino acid pools, J. T. Holden, Ed., pp. 73–108. Amsterdam-London-New York: Elsevier 1962
Indge, K. J.: Metabolic lysis of yeast protoplasts. J. gen. Microbiol. 51, 433–440 (1968a)
Indge, K. J.: Polyphosphates of the yeast vacuole. J. gen. Microbiol. 51, 447–455 (1968b)
Kotyk, A., Rihova, L.: Transport of α-aminoisobutyric acid in Saccharomyces cerevisiae. Feedback control. Biochim. biophys. Acta (Amst.) 288, 380–389 (1972)
Ling, G. N.: A new model for the living cell: a summary of the theory and recent experimental evidence in its support. Int. Rev. Cytol. 26, 1–62 (1969)
Matile, Ph., Wiemken, A.: The vacuole as the lysosome of the yeast cell. Arch. Mikrobiol. 56, 148–155 (1967)
Meyenburg, H. K. von: The budding cycle of Saccharomyces cerevisiae. Path. et Microbiol. 31, 117–127 (1968)
Middelhoven, W. J.: Induction and repression of arginase and ornithine transaminase in baker's yeast. Antonie v. Leeuwenhoek 36, 1–19 (1970)
Nurse, P., Wiemken, A.: Amino acid pools and metabolism during the cell division cycle of arginine-grown Candida utilis. J. Bact. 117, 1108–1116 (1974)
Oaks, A., Bidwell, R. G. S.: Compartmentation of intermediary metabolites. Ann. Rev. Plant Physiol. 21, 43–66 (1970)
Ramos, F., Thuriaux, P., Wiame, J. M., Bechet, J.: The participation of ornithine and citrulline in the regulation of arginine metabolism in Saccharomyces cerevisiae. Europ. J. Biochem. 12, 40–47 (1970)
Roush, A. H.: Crystallization of purines in the vacuole of Candida utilis. Nature (Lond.) 190, 449 (1961)
Ryan, E. D., Tracy, J. W., Kohlhaw, G. B.: Subcellular localization of the leucine biosynthetic enzymes in yeast. J. Bact. 116, 222–225 (1973)
Sims, A. P., Folkes, B. F.: A kinetic study of the assimilation of [15N]ammonia and the synthesis of amino acids in an exponentially growing culture of Candida utilis. Proc. roy. Soc. B 159, 479–502 (1964)
Sols, A., Marco, R.: Concentrations of metabolites and binding sites. Implications in metabolic regulation. In: Current topics in cellular regulation, Vol. 2, B. L. Horecker and E. R. Stadtman, Eds., pp. 227–273. New York: Academic Press 1970
Stebbing, N.: Precursor pools and endogenous control of enzyme synthesis in biosynthetic pathways. Bact. Rev. 38, 1–28 (1974)
Subramanian, K. N., Weiss, R. L., Davis, R. H.: Use of external, biosynthetic, and organellar arginine by Neurospora. J. Bact. 115, 284–290 (1973)
Svihla, G., Dainko, J. C., Schlenk, F.: Ultraviolet microscopy of purine compounds in the yeast vacuole. J. Bact. 83, 399–402 (1963)
Weiss, R. L.: Intracellular localization of ornithine and arginine pools in Neurospora. J. Biol. Chem. 248, 5401–5413 (1973)
Weiss, R. L., Davis, R. H.: Intracellular localization of enzymes of arginine metabolism in Neurospora. J. biol. Chem. 248, 5403–5408 (1973)
Wiemken, A.: Eigenschaften der Hefevakuole. Thesis Nr. 4340, Swiss Federal Institute Technol., Zürich 1969
Wiemken, A., Matile, P., Moor, H.: Vacuolar dynamics in synchronously budding yeast. Arch. Mikrobiol. 70, 89–103 (1970)
Wiemken, A., Nurse, P.: Isolation and characterization of the amino acids pools located within the cytoplasm and vacuoles of Candida utilis. Plant (Berl.) 109, 293–306 (1973)
Witt, I., Holzer, H.: Hauptweg des NH4 +-Einbaues in Glucose-oxydierender Bäckerhefe. Biochem. Z. 339, 255–265 (1964)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wiemken, A., Dürr, M. Characterization of amino acid pools in the vacuolar compartment of Saccharomyces cerevisiae . Arch. Microbiol. 101, 45–57 (1974). https://doi.org/10.1007/BF00455924
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00455924