Abstract
The energy-dependent transport of phenylalanine into isolated vacuoles of barley (Hordeum vulgare L.) mesophyll protoplasts has been studied by silicone-layer floatation filtering. The uptake of this aromatic amino acid into the vacuolar compartment is markedly increased by MgATP, showing saturation kinetics; the K m values were 0.5 mM for MgATP and 1.2 mM for phenylalanine. V max for phenylalanine transport was estimated to 140 nmol phenylalanine·(mg·Chl)-1·h-1. The transport shows a distinct pH optimum at 7.3 and is markedly inhibited by 40 mM nitrate. Azide (1 mM) and vanadate (400 μM) had no or little effect on rates of transport while p-fluorophenylalanine seemed to be an effective inhibitor, indicating a possible competition at an amino-acid carrier. Ionophores such as valinomycin, nigericin or gramicidin were strong inhibitors of phenylalanine transport, indicating that this process is coupled to both the transmembrane pH gradient (ΔpH) and the transmembrane potential (ΔΨ).
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Abbreviations
- BSA:
-
bovine serum albumin
- Chl:
-
chlorophyll
- Hepes:
-
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- ΔpH:
-
transmembrane pH gradient
- ΔΨ:
-
transmembrane potential
References
Arnon, D.I. (1949) Copper enzymes in isolated chloroplasts. Polyphenol oxidase in Beta vulgaris. Plant Physiol. 24, 1–15
Asami, S., Hara-Nishimura, I., Nishimura, M., Akazawa, T. (1985) Translocation of photosynthates into vacuoles in spinach leaf protoplasts. Plant Physiol. 77, 963–968
Boller, T., Kende, H. (1979) Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 63, 1123–1132
Driessen, A.J.M., Hellingwerf, K.J., Konings, W.N. (1987) Mechanism of energy coupling to entry and exit of neutral and branched chain amino acids in membrane vesicles of Streptococcus cremoris. J. Biol. Chem. 262, 12438–12443
Fiedler, E., Schultz, G. (1985) Localization, purification, and characterization of shikimate-oxidoreductase-dehydroquinate hydrolyase from stroma of spinach chloroplasts. Plant Physiol. 79, 212–218
Heber, U., Pon, N.G., Heber, M. (1963) Localization of carboxydismutase and triosephosphate dehydrogenases in chloroplasts. Plant Physiol. 38, 355–360
Heldt, H.W., Flügge, U.I. (1986) Intrazellulärer Transport in grünen Pflanzenzellen. Naturwissenschaften 73, 1–7
Jung, K.-D., Lüttge, U. (1980) Amino acid uptake by Lemna gibba by a mechanism with affinity to neutral L-and D-amino acids. Planta 150, 230–235
Jung, K.-D., Lüttge, U. (1982) Inhibition of the amino acid co-transport of Lemna gibba by incorporation of p-fluorophenylalanine. Physiol. Plant. 55, 231–235
Jung, K.-D., Lüttge, U. Fischer, E. (1982) Uptake of neutral and acidic amino acids by Lemna gibba correlated with the H+-electrochemical gradient at the plasmalemma. Physiol. Plant. 55, 351–355
Kaiser, G., Martinoia, E., Schmidt, J.M., Hincha, D.K., Heber, U. (1986) Polypeptide pattern and enzymatic character of vacuoles isolated from barley mesophyll protoplasts. Planta 169, 345–355
Kaiser, G., Martinoia, E., Wiemken, A. (1982) Rapid appearance of photosynthetic products in the vacuoles isolated from barley mesophyll protoplasts by a new fast method. Z. Pflanzenphysiol. 107, 103–113
Kelly, G.J., Gibbs, M. (1973) Nonreversible D-glyceraldehyde-3-phosphate dehydrogenase of plant tissues. Plant Physiol. 53, 111–118
Leuschner, C., Homeyer, U., Schultz, G. (1988) Chloroplasts control the rates of aromatic as well as of branched chain amino acid synthesis in Hordeum (Abstr.) Plant Physiol. 86 Suppl., S 877
Martinoia, E., Flügge U.I., Kaiser, G., Heber, U., Heldt, H.W. (1985) Energy-dependent uptake of malete into vacuoles isolated from barley mesophyll protoplasts. Biochim. Biophys. Acta 806, 311–319
Nishida, K., Tominaga, O. (1987) Energy-dependent uptake of malate into vacuoles from CAM plant, Kalanchoe daigremontiana. J. Plant Physiol. 127, 385–393
Schulze-Siebert, D., Homeyer, U., Soll, J., Schultz, G. (1987) Synthesis of plastoquinone-9, α-tocopherol and phylloquinone (vitamin K1) and its integration in chloroplast carbon metabolism of higher plants. In: The metabolism, structure, and function of plant lipids, pp. 29–36, Stumpf, P.K., Mudd, J.B., Nes, W.D. eds. Plenum, New York
Stitt, M., Bulpin, P.V., ap Rees, T. (1978) Pathway of starch breakdown in photosynthetic tissues of Pisum sativum. Biochim. Biophys. Acta 544, 200–214
Stitt, M., McC. Lilley, R., Heldt, H.W. (1982) Adenine nucleotide levels in the cytosol. chloroplasts, and mitochondria of wheat leaf protoplasts. Plant Physiol. 70, 971–977
Sze, H. (1985) H+-translocating ATPases: advances using membrane vesicles. Annu. Rev. Plant Physiol. 36, 175–208
Tolbert N.E., Yamazaki, R.K., Oeser, H. (1970) Localization and properties of hydroxypyruvate and glyoxylate reductase in spinach leaf particles. J. Biol. Chem. 245, 5129–5136
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Homeyer, U., Schultz, G. Transport of phenylalanine into vacuoles isolated from barley mesophyll protoplasts. Planta 176, 378–382 (1988). https://doi.org/10.1007/BF00395418
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DOI: https://doi.org/10.1007/BF00395418