Abstract
The localization of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase was examined in suspension-cultured cells of tomato (Lycopersicon esculentum Mill.), using cell-fractionation techniques, followed by immunoblot analysis with monospecific antibodies raised against a tomato ACC oxidase expressed in Escherichia coli. When assayed in vivo, ACC oxidase had a low activity in untreated tomato cells but was strongly induced when the cells were supplied with its substrate, ACC. Immunoblots showed that this induction was accompanied by the accumulation of a single protein corresponding to ACC oxidase, with an apparent molecular mass (Mr) of 36 kDa. The level of this protein in induced cells, estimated by immunoblotting, was compared with that in protoplasts and vacuoles, and with that in various particulate and soluble fractions obtained by differential centrifugation of cell homogenates. It was found that the ACC oxidase antigen was absent from the vacuole, and that most of it was localized in the cytoplasm of the protoplasts without being associated with membranes. Measurements of ACC oxidase activity in preparations of protoplasts and vacuoles supported these results.
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Abbreviations
- ACC:
-
1-aminocyclopropane-1-carboxylic acid
References
Bergmeyer, H.U. (1974) Methoden der enzymatischen Analyse, vol. 1. Verlag Chemie, Weinheim
Boller, T., Kende, H. (1979) Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 63, 1123–1132
Bouzayen, M., Latché, A., Pech, J.-C. (1990) Subcellular localization of the sites of conversion of 1-aminocyclopropane-1-carboxylic acid into ethylene in plant cells. Planta 180, 175–180
Bradley, D.J., Kjellbom, P., Lamb, C.J. (1992) Elicitor and wound induced oxidative cross-linking of a proline-rich plant cell wall protein: A novel, rapid defense response. Cell 70, 21–30
Erdmann, H., Griesbach, R.J., Lawson, R.H., Mattoo, A.K. (1989) 1-Aminocyclopropane-1-carboxylic-acid-dependent ethylene production during re-formation of vacuoles in evacuolated pro toplasts of Petunia hybrida. Planta 179, 196–202
Felix, G., Grosskopf, D.G., Regenass, M., Basse, C.W., Boller, T. (1991) Elicitor-induced ethylene biosynthesis in tomato cells. Characterization and use as a bioassay for elicitor action. Plant Physiol. 97, 19–25
Guy, M. (1990) Vacuolar ethylene formation does not depend on membrane potential. Physiol. Plant. 79, 526–530
Guy, M., Kende, H. (1984) Conversion of 1-aminocyclopropane-1-carboxylic acid to ethylene by isolated vacuoles of Pisum sativum L. Planta 160, 281–287
Hamilton, A.J., Bouzayen, M., Grierson, D. (1991) Identification of a tomato gene for the ethylene-forming enzyme by expression in yeast. Proc. Natl. Acad. Sci. USA 88, 7434–7437
Harlow, E., Lane, D. (1988) Antibodies: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Hochuli, E., Bannwarth, W., Döbeli, H., Gentz, R., Stüber, D. (1988) Genetic approach to facilitate purification of recombinant proteins with a novel motal chelate adsorbent. Bio/Technology 6, 1321–1325
Kende, H. (1993) Ethylene biosynthesis. Annu. Rev. Plant Physiol. Plant Mol. Biol. 44, 283–307
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
Mayne, R.G., Kende, H. (1986) Ethylene biosynthesis in isolated vacuoles of Vicia faba L. — requirement for membrane integrity. Planta 167, 159–165
Mattoo, A.K., Lieberman, M. (1977) Localization of the ethylene-synthesizing system in apple tissue. Plant Physiol. 60, 794–799
Peck, S.C., Reinhardt, D., Olson, D.C., Boller, T., Kende, H. (1992) Localization of the ethylene-forming enzyme from tomatoes, 1-aminocyclopropane-1-carboxylic acid oxidase, in transgenic yeast. J. Plant Physiol. 140, 681–686
Porter, A.J.R., Borlakoglu, J.T., John, P. (1986) Activity of the ethylene-forming enzyme in relation to plant cell structure and organisation. J. Plant Physiol. 125, 207–216
Rombaldi, C., Lelièvre, J.-M., Latché, A., Petitprez, M., Bouzayen, M., Pech, J.-C. (1994) Immunocytolocalization of 1-aminocyclo-propane-1-carboxylic acid oxidase in tomato and apple fruit. Planta 192, 453–460
Spanu, P., Reinhardt, D., Boller, T. (1991) Analysis and cloning of the ethylene-forming enzyme from tomato by functional expression of its mRNA in Xenopus laevis oocytes. EMBO J. 10, 2007–2013
Ververidis, P., John, P. (1991) Complete recovery in vitro of ethylene forming enzyme activity. Phytochemistry 30, 725–727
Vögeli-Lange, R., Wagner, G.J. (1990) Subcellular localization of cadmium and cadmium-binding peptides in tobacco leaves. Plant Physiol. 92, 1086–1093
Yang, S.F., Huffman, N.E. (1984) Ethylene biosynthesis and its regulation in higher plants. Annu Rev. Plant Physiol. 35, 155–189
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We thank Martin Regenass (Friedrich Miescher-Institut, Basel, Switzerland) for maintaining the cell cultures and Georg Felix (Friedrich Miescher-Institut, Basel, Switzerland) for helpful discussions. This work was supported, in part, by the Swiss National Science Foundation, Grant 31-26492.89.
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Reinhardt, D., Kende, H. & Boiler, T. Subcellular localization of 1-aminocyclopropane-1-carboxylate oxidase in tomato cells. Planta 195, 142–146 (1994). https://doi.org/10.1007/BF00206302
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DOI: https://doi.org/10.1007/BF00206302