Summary
An artificially produced electrochemical potential difference for protons (protonmotive force) provided the energy for the transport of galactosides inEscherichia coli cells which were depleted of their endogenous energy reserves. The driving force for the entry of protons was provided by either a transmembrane pH gradient or a membrane potential. The pH gradient across the membrane was created by acidifying the external medium. The membrane potential (inside negative) was established by the outward diffusion of potassium (in the presence of valinomycin) or by the inward diffusion of the permeant thiocyanate ion. The magnitude of the electrochemical potential difference for protons agreed well with magnitude of the chemical potential difference of the lactose analog, thiomethylgalactoside. The observations are consistent with the view that the carrier-mediated entry of each galactoside molecule is accompanied by the entry of one proton.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altendorf, K., Harold, F. M., Simoni, R. D. 1974. Impairment and restoration of the energized state in membrane vesicles of a mutant ofEscherichia coli lacking adenosine triphosphatase.J. Biol. Chem. 249:4587
Berger, E. A. 1973. Different mechanisms of energy coupling for the active transport of proline and glutamine inEscherichia coli.Proc. Nat. Acad. Sci. USA 70:1514
Berger, E. A., Heppel, L. A. 1974. Different mechanisms of energy coupling for the shock-sensitive and shock-resistant amino acid permeases ofEscherichia coli.J. Biol. Chem. 249:7747
Butlin, J. D., Cox, G. B., Gibson, F. 1971. Oxidative phosphorylation inEscherichia coli K12: Mutations affecting magnesium ion- or calcium ion-stimulated adenosine triphosphatase.Biochem. J. 124:75
Cohen, G. N., Rickenberg, H. V. 1956. Concentration specifique reversible des amino acides chezEscherichia coli.Ann. Inst. Pasteur, Paris 91:693
Davila, H. V., Salzberg, B. M., Cohen, L. B., Waggoner, A. S. 1973. A large change in axon fluorescence that provides a promising method for measuring a membrane potential.Nature New Biol. 241:159
Flagg, J. L., Wilson, T. H. 1976. Galactoside accumulation byEscherichia coli. driven by a pH gradient.J. Bacteriol. 125:1235
Fox, C. F., Wilson, G. 1968. The role of a phosphoenolpyruvate-dependent kinase system in β-glucoside catabolism inEscherichia coli.Proc. Nat. Acad. Sci. USA 59:988
Grinius, L., Slušnyte, R., Griniuviené, B. 1975. ATP synthesis driven by protonmotive force imposed acrossEscherichia coli cell membranes.FEBS Lett. 57:290
Griniuviené, B., Chmieliauskaite, V., Melvydas, V., Dzhega, P., Grinius, L. 1975. Conversion ofEscherichia coli cell-produced metabolic energy into electric form.Bioenergetics 7:17
Harold, F. M., Altendorf, K. 1974. Cation transport in bacteria: K+, Na+ and H+.In: Current Topics in Membranes and Transport. F. Bronner and A. A. Kleinzeller, editors. Vol. 5., p. 1. Academic Press, New York
Harold, F. M., Pavlasova, E., Baarda, J. R. 1970. A transmembrane pH gradient inStreptococcus faecalis: Origin, and dissipation by proton conductors and N, N′-dicyclohexylcarbodiimide.Biochim. Biophys. Acta 196:235
Hirata, H., Altendorf, K., Harold, F. M. 1973. Role of an electrical potential in the coupling of metabolic energy to active transport by membrane vesicles ofEscherichia coli.Proc. Nat. Acad. Sci. USA 70:1804
Hirata, H., Altendorf, K., Harold, F. M. 1974. Energy coupling in membrane vesicles ofEscherichia coli. I. Accumulation of metabolites in response to an electrical potential.J. Biol. Chem. 249:2939
Hoffman, J. F., Laris, P. C. 1974. Determination of membrane potentials in human andamphiuma red blood cells by means of a fluorescent probe.J. Physiol. 239:519
Kaback, H. R. 1971. Bacterial membranes.In: Methods in Enzymology. W. B. Jakoby, editor. Vol. 22, p. 99. Academic Press, New York
Kashket, E. R., Wilson, T. H. 1972. Galactoside accumulation associated with ion movements inStreptococcus lactis.Biochem. Biophys. Res. Commun. 49:615
Kashket, E. R., Wilson, T. H. 1973. Proton-coupled accumulation of galactoside inStreptococcus lactis 7962.Proc. Nat. Acad. Sci. USA 79:2866
Kashket, E. R., Wilson, T. H. 1974. Protonmotive force in fermentingStreptococcus lactis 7962 in relation to sugar accumulation.Biochem. Biophys. Res. Commun. 59:879
Kashket, E. P., Wong, P. T. S. 1969. The intracellular pH ofEscherichia coli.Biochim. Biophys. Acta 193:212
Larris, P. C., Pershadsingh, H. A. 1974. Estimations of membrane potentials inStreptococcus faecalis by means of a fluorescent probe.Biochem. Biophys. Res. Commun. 57:620
Maloney, P. C., Kashket, E. R., Wilson, T. H. 1975. A protonmotive force drives ATP synthesis in bacteria.Proc. Nat. Acad. Sci. USA 71:3896
Maloney, P. C., Kashket, E. R., Wilson, T. H. 1975. Methods for studying transport in bacteria.In: Methods in Membrane Biology. E. D. Korn, editor. Vol. 5, p. 1. Plenum Press, New York
Maloney, P. C., Wilson, T. H. 1975. ATP synthesis driven by a protonmotive force inStreptococcus lactis.J. Membrane Biol. 25:285
Mitchell, P. 1963. Molecule, group and electron translocation through natural membranes.Biochem. Soc. Symp. 22:142
Pavlasova, E., Harold, F. M. 1968. Energy coupling in the transport of β-galactosides byEscherichia coli: Effect of proton conductors.J. Bacteriol. 98:198
Roos, A. 1965. Intracellular pH and intracellular buffering power of the cat brain.Am. J. Physiol. 209:1233
Sâsárman, A., Surdeanu, M., Horodniceanu, T. 1968. Locus determining the synthesis of γ-aminolevulinic acid inEscherichia coli K-12.J. Bacteriol. 96:1882
Schuldiner, S., Kaback, H. R. 1975. Membrane potential and active transport in membrane vesicles fromEscherichia coli.Biochemistry 14:5451
Simoni, R. D., Postma, P. W. 1975. The energetics of bacterial active transport.Annu. Rev. Biochem. 44:523
Simoni, R. D., Shallenberger, M. K. 1972. Coupling of energy to active transport of amino acids inEscherichia coli.Proc. Nat. Acad. Sci. USA 69:2663
Singh, A. P., Bragg, P. D. 1974. Energization of phenylalanine transport and energy-dependent transhydrogenase by ATP in cytochrome-deficientEscherichia coli.Biochem. Biophys. Res. Commun. 41:655
Tsuchiya, T., Rosen, B. P. 1976. ATP synthesis by an artificial proton gradient in right-side-out membrane vesicles ofEscherichia coli.Biochem. Biophys. Res. Commun. 68:497
West, I. C. 1970. Lactose transport coupled to proton movements inEscherichia coli.Biochem. Biophys. Res. Commun. 41:655
West, I. C., Mitchell, P. 1972. Proton coupled β-galactoside translocation in non-metabolizingEscherichia coli.J. Bioenergetics. 3:445
West, I. C., Mitchell, P. 1973. Stoichiometry of lactose-H+ symport across the plasma membrane ofEscherichia coli.Biochem. J. 132:587
Wilson, D. M., Alderete, J. F., Maloney, P. C., Wilson, T. H. 1976. A protonmotive force as the source of energy for adenosine 5′-triphosphate synthesis inEscherichia coli.J. Bacteriol. 126:327
Winkler, H. H., Wilson, T. H. 1966. The role of energy coupling in the transport of β-galactosides byEscherichia coli.J. Biol. Chem. 241:2200
Wood, J. M. 1975. Leucine transport inEscherichia coli: The resolution of multiple transport systems and their coupling to metabolic energy.J. Biol. Chem. 250:4477
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Flagg, J.L., Wilson, T.H. A protonmotive force as the source of energy for galactoside transport in energy depletedEscherichia Coli . J. Membrain Biol. 31, 233–255 (1977). https://doi.org/10.1007/BF01869407
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01869407