Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species

THUME M, Dietz K-J (1991)
Planta 185(4): 569-575.

Journal Article | Published | English

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The tonoplast amino-acid transporter of barley (Hordeum vulgare L.) mesophyll cells was functionally reconstituted by incorporating solubilized tonoplast membranes, vacuoplast membranes or tonoplast-enriched microsomal vesicles into phosphatidylcholine liposomes. (i) Time-, concentration- and ATP-dependence of amino-acid uptake were similar to results with isolated vacuoles. Although the orientation of incorporation could not be controlled, the results indicate that the transporter functions as a uniport system which allows regulated equilibration by diffusion between the cytosolic and vacuolar amino-acid pools. (ii) The ATP-modulated amino-acid carrier was also successfully reconstituted from barley epidermal protoplasts and Valerianella or Tulipa vacuoplasts, indicating its general occurrence. (iii) Fractionation of solubilized tonoplasts by size-exclusion chromatography followed by reconstitution of the fractions for glutamine transport gave two activity peaks: the first eluted in the region of high-molecular-mass vesicles and the second at a size of 300 kDa for the Triton-protein micelle.
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THUME M, Dietz K-J. Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species. Planta. 1991;185(4):569-575.
THUME, M., & Dietz, K. - J. (1991). Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species. Planta, 185(4), 569-575.
THUME, M., and Dietz, K. - J. (1991). Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species. Planta 185, 569-575.
THUME, M., & Dietz, K.-J., 1991. Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species. Planta, 185(4), p 569-575.
M. THUME and K.-J. Dietz, “Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species”, Planta, vol. 185, 1991, pp. 569-575.
THUME, M., Dietz, K.-J.: Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species. Planta. 185, 569-575 (1991).
THUME, M, and Dietz, Karl-Josef. “Reconstitution of the tonoplast amino acid carrier into liposomes. Evidence for an ATP regulated carrier in different species”. Planta 185.4 (1991): 569-575.
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PMID: 24186536
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