Abstract
A new procedure for the isolation of membrane vesicles fromAcholeplasma laidlawii cells is described. The membrane vesicles are completely free from contaminations of whole cells and cell debris and represent a homogeneous fraction as shown by electron microscopy, Ficoll density-gradient centrifugation, and titration on agar plate. Absence of cytoplasmic contaminations was confirmed by double-labelling of membranes with3H-oleic acid and14C-uridine, as well as by distribution of specific marker enzymes of membranes and cytoplasm. On the basis of light-scattering and electron microscopy, the vesicular nature of these membranes was established. The vesicles had the same orientation as intact cells (absence on membrane vesicles of ATPase and NADH dehydrogenase activities, localized in the inner surface of membrane). The respiratory activity of the membrane vesicles was low and was not stimulated by exogenous substrates, the respiratory chain of the vesicles being reduced and terminated by flavoproteins. The ability of membrane vesicles to take up carbohydrates was shown.
Similar content being viewed by others
References
Altendorf K., Staehelin L.: Orientation of membrane vesicles fromEscherichia coli as detected by freeze-cleave electron microscopy.J. Bacteriol. 117, 888 (1974).
Anderson R. L., Kamel M. G.: Hexokinase, p. 388, inMethods in Enzymology (Wood W. A., ed.), Vol. 9, Academic Press Inc. New York, 1966.
Asano A., Cohen S. N., Baker R. G., Brodie A. F.: Orientation of the cell membrane in ghost and electron transport particles ofMycobacterium phlei.J. Biol. Chem. 248, 3386 (1973).
De Kruyff B., Demel R. A., Van Deenen L. L. M.: The effect of cholesterol and epicholesterol incorporation on the permeability and on the phase transition of intactAcholeplasma laidlawii cell membranes and derived liposomes.Biochim. Biophys. Acta 255, 331 (1972).
Evtodienko Y. V., Mokchova E. N.: Quantitative determination of cytochrome correlations in mitochondria, p. 35, inThe Mechanisms of Respiration, Photosynthesis and Nitrogen Fixation. Nauka, Moscow, 1967.
Fedotov N. S., Panchenko L. F., Tarshis M. A.: Isolation of membrane preparations fromAcholeplasma laidlawii cells and their transport characteristics. (In Russian)Microbiologiya 43, 543 (1974).
Futai M.: Orientation of membrane vesicles fromE. coli prepared by different procedures.J. Membrane Biol. 15, 15 (1974).
Gershanovich V. N.: Biocatalysts performing carbohydrate transport in bacteria. (In Russian)Usp. sovremen. biol. 72, 24 (1971).
Gordon A. S., Lombardi F. J., Kaback H. R.: Solubilization and partial purification of amino acid-specific components on thed-lactate dehydrogenase-coupled amino acid-transport system.Proc. Nat. Acad. Sci. USA 69, 358 (1972).
Gorneva G. H., Ryabova I. D.: Membrane orientation in vesicles fromM. lysodeikticus cells.FEBS Lett. 42, 271 (1974).
Kaback H. P.: Transport across isolated bacterial cytoplasmic membranes.Biochim. Biophys. Acta 265, 367 (1972).
Kahane S., Razin S.: pH-dependent changes in density of plasma membranes of growingMycoplasma laidlawii cells.FEBS Lett. 10, 261 (1970).
Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J.: Protein measurement with the Folin phenol reagent.J. Biol. Chem. 193, 265 (1951).
Maniloff J.: Ultrastructure ofM. laidlawii during culture development.J. Bacteriol. 102, 561 (1970).
McElhaney R. N., de Gier J., van der Neut-Kok E. C. M.: The effect of alteration in fatty acid composition and cholesterol content on the nonelectrolyte permeability ofAcholeplasma laidlawii B cells and derived liposomes.Biochim. Biophys. Acta 298, 500 (1973).
Ng F. M.-W., Dames E. A.: Chemostat studies on the regulation of glucose metabolism inPseudomonas aeruginosa by citrate.Biochem. J. 132, 129 (1973).
Pollack Y. D., Razin S., Cleverdon R. C.: Localization of enzymes inMycoplasma.J. Bacteriol. 90, 617 (1965).
Razin S.: The Mycoplasma membranes, p. 317 inThe Mycoplasmatales and the L-phase of Bacteria (Hayflick, L., ed.), Appelton-Century-Crofts, New York, 1969.
Rottem S., Cirillo V. P., de Kruyff B., Schinitzky M., Razin S.: Cholesterol in Mycoplasma membranes.Biochim. Biophys. Acta 323, 509 (1973).
Tarshis M. A., Migoushina V. L., Panchenko L. F., Fedotov N. S., Bourd H. I.: Studies of sugar transport inA. laidlawii cells.Eur. J. Biochem. 40, 171 (1973).
Van der Neut-Kok E. C. M., de Gier J., Middelbeek E. J., Van Deenen L. L. M.: Valinomycin-induced potassium and rubidium permeability of intact cells ofAcholeplasma laidlawii B.Biochim. Biophys. Acta 332, 97 (1974).
Van Thienen G., Postma P. W.: Coupling between energy conservation and active transport of serine inEscherichia coli.Biochim. Biophys. Acta 323, 429 (1973).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fedotov, N.S., Panchenko, L.F., Logachev, A.P. et al. Transport properties of membrane vesicles fromAcholeplasma laidlawii . Folia Microbiol 20, 470–479 (1975). https://doi.org/10.1007/BF02891706
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02891706