Energetics of alanine, lysine, and proline transport in cytoplasmic membranes of the polyphosphate-accumulating Acinetobacter johnsonii strain 210A

H. W. Van Veen*, T. Abee, A. W. F. Kleefsman, B. Melgers, G. J. J. Kortstee, W. N. Konings, A. J. B. Zehnder

*Corresponding author for this work

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    Abstract

    Amino acid transport in right-side-out membrane vesicles of Acinetobacter johnsonii 210A was studied. L-Alanine, L-lysine, and L-proline were actively transported when a proton motive force of -76 mV tvas generated by the oxidation of glucose via the membrane-bound glucose dehydrogenase. Kinetic analysis of amino acid uptake at concentrations of up to 80 mu M revealed the presence of a single transport system for each of these amino acids with a K-t of less than 4 mu M. The mode of energy coupling to solute uptake was analyzed by imposition of artificial ion diffusion gradients. The uptake of alanine and lysine was driven by a membrane potential and a transmembrane pH gradient. In contrast, the uptake of proline was driven by a membrane potential and a transmembrane chemical gradient of sodium ions. The mechanistic stoichiometry for the solute and the coupling ion was close to unity for all three amino acids. The Na+ dependence of the proline carrier was studied in greater detail. Membrane potential-driven uptake of proline was stimulated by Na+, with a half-maximal Na+ concentration of 26 mu M. At Na+ concentrations above 250 mu M, proline uptake was strongly inhibited. Generation of a sodium motive force and maintenance of a low internal Na+ concentration are most likely mediated by a sodium/proton antiporter, the presence of which was suggested by the Na+-dependent alkalinization of the intravesicular pH in inside-out membrane vesicles. The results show that both H+ and Na+ can function as coupling ions in amino acid transport in Acinetobacter spp.

    Original languageEnglish
    Pages (from-to)2670-2676
    Number of pages7
    JournalJournal of Bacteriology
    Volume176
    Issue number9
    DOIs
    Publication statusPublished - May-1994

    Keywords

    • AMINO-ACID-TRANSPORT
    • ESCHERICHIA-COLI
    • SALMONELLA-TYPHIMURIUM
    • STREPTOCOCCUS-CREMORIS
    • LACTOCOCCUS-LACTIS
    • BACTERIAL-CELLS
    • VESICLES
    • PH
    • RECONSTITUTION
    • DEHYDROGENASE

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