Abstract
Highly purified tonoplast fractions isolated by preparative free-flow electrophoresis from hypocotyls of etiolated soybean (Glycine max L. (Merr.)) were examined by negative-staining electron microscopy, and many but not all vesicles were found to exhibit head and stalk structures resembling the 9-nm stalked F1 ATPase particles reported previously for Neurospora (Bowman et al., 1989, J. Biol. Chem. 264, 15606–15612). The structures show distinguishing characteristics similar to those for Neurospora. These include a cleft in the particle not exhibited by mitochondrial F1 ATPase and a tendency to disappear from the membrane when treated with nitrate plus Mg−2+-ATP-containing solutions. The position of the stalked ATPase structures, indicates that some of the tonoplast vesicles were oriented cytoplasmic side out whereas others were oriented cytoplasmic side in.
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Adachi, I., Arai, H., Pimental, R., Forgac, M. (1990a) Proteolysis and orientation on reconstitution of the coated vesicle proton pump. J. Biol. Chem. 265, 960–966
Adachi, I., Puopolo, K., Marquez-Sterling, N., Arai, H., Forgac, M. (1990b) Dissociation, cross-linking, and glycosylation of the coated vesicle pump. J. Biol. Chem. 265, 967–973
Al-Aquati, Q. (1986) Proton-translocating ATPases. Annu. Rev. Cell Biol. 2, 179–199
Arai, H., Berne, M., Terres, G., Terres, H., Puopolo, K., Forgac, M. (1987) Subunit composition and ATP site. Labeling of the coated vesicle proton-translocating adenosinetriphosphatase. Biochemistry 26, 6632–6638
Bowman, B.J., Bowman, E.J. (1986) H−+-ATPases from mitochondria, plasma membranes and vacuoles of fungal cells. J. Membr. Biol. 94, 83–97
Bowman, B.J., Dschida, W.J., Harris, T., Bowman, E.J. (1989) The vacuolar ATPase of Neurospora crassa contains an F1-like structure. J. Biol. Chem. 264, 15606–15612
Bowman, E.J., Mandala, S., Taiz, L., Bowman, B.J. (1986) Structural studies of the vacuolar membrane ATPase from Neurospora crassa and comparisons with the tonoplast membrane ATPase from Zea mays. Proc. Natl. Acad. Sci. USA 83, 48–52
Bradford, M.M. (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254
Canut, H., Brightman, A.O., Boudet, A.M., Morré, D.J. (1987) Determination of sidedness of plasma membrane and tonoplast vesicles isolated from plant stems. In: Plant membranes. Structure, function, biogenesis, pp. 141–159, Leaver, C., Sze, H., eds. Alan R. Liss, New York
Forgac, M. (1989) Structure and function of vacuolar class of ATP-driven proton pumps. Physiol. Rev. 69, 765–796
Hirata, R., Ohsumi, Y., Nakano, A., Kawasaki, H., Suzuki, K., Anraku, Y. (1990) Molecular structure of a gene, VMA1, encoding the catalytic subunit of H−+-translocating adenosine triphosphatase from vacuolar membranes of Saccharomyces cerevisiae. J. Biol. Chem. 265, 6726–6733
Lai, S., Randall, S.K., Sze, H. (1988) Peripheral and integral subunits of the tonoplast H−+-ATPase from oat roots. J. Biol. Chem. 263, 16731–16737
Lichko, L.P., Okorokov, L.A. (1985) What family of ATPases does the vacuolar H−+-ATPase belong to? FEBS Lett. 187, 349–353
Mandala, S., Taiz, L. (1985) Proton transport in isolated vacuoles from corn coleoptiles. Plant Physiol. 78, 327–333
Manolson, M.F., Rea, P.A., Poole, R.J. (1985) Identification of 3-O-(4-benzoyl)benzoyladenosine S−1-triphosphateand N,N−1-dicylohexylcarbodiimide-binding subunits of a higher plant H−+-translocating ATPase. J. Biol. Chem. 260, 12273–12279
Mellman, I., Fuchs, R., Helenius, A. (1986) Acidification of the endocytic and exocytic pathways. Annu. Rev. Biochem. 55, 663–700
Moriyama, Y., Nelson, N. (1987) The purified ATPase from chromaffin granule membranes is an anion-dependent proton pump. J. Biol. Chem. 262, 9175–9180
Moriyama, Y., Nelson, N. (1989) Cold inactivation of vacuolar proton-ATPases. J. Biol. Chem. 264, 3577–3582
Morré, D.J., Auderset, G., Penel, C., Canut, H. (1987) Cytochemical localization of NADH-ferricyanide oxido-reductase in hypocotyl segments and isolated membrane vesicles of soybean. Protoplasma 140, 133–140
Parry, R.V., Turner, J.C., Rea, P.A. (1989) High purity preparation of higher plant vacuolar H−+-ATPasereveal additional subunits. Revised subunit composition. J. Biol. Chem. 264, 20025–20032
Penefsky, H.S., Pullman, M.E., Datta, A., Racker, E. (1960) Partial resolution of the enzymes catalyzing oxidative phosphorylation II. Participation of a soluble adenosine triphosphatase in oxidative phosphorylation. J. Biol. Chem. 35, 3330–3336
Percy, J.M., Pryde, J.G., Apps, D.K. (1985) Isolation of ATPase I, the protein pump of chromaffin granule membrane. Biochem. J. 231, 57–564
Randall, S.K., Sze, H. (1986) Properties of the partially purified tonoplast H−+-pumping ATPase from oat roots. J. Biol. Chem. 262, 1364–1371
Sandelius, A.S., Penel, C., Auderset, G., Brightman, A., Millard, M., Morré, D.J. (1986) Isolation of highly purified fractions of membrane and tonoplast from the same homogenate of soybean hypocotyls by free-flow electrophoresis. Plant Physiol. 81, 177–185
Scherer, G.F.E. (1984) Subcellular localization of H−+-ATPase from pumpkin hypocotyls. (Cucurbita maxima L.) by membrane fractionation. Planta 160, 348–356
Smith, P.K., Krohn, R.I., Hermanson, G.T., Mallia, A.K., Gartner, F.H., Provenzano, M.D., Fujimoto., E.K., Goeke, N.M., Olson, B.J., Klenk, D.C. (1985) Measurement of protein using bicinchoninic acid. Anal. Biochem. 150, 76–85
Sze, H. (1985) H−+-translocating ATPases: Advances using membrane vesicles. Annu. Rev. Plant Physiol. 36, 175–208
Taiz, L., Gogarten, J.P., Kibak, H., Struve, I., Bernasconi, P., Rausch, T., Taiz, S.L. (1989) Studies on the structure and evolution of vacuolar H−+-ATPase. In: Plant membrane transport: the current position, pp. 131–137, Dainty, J., de Michelis, M.C., Marré, E., Rasi-Caldogno, F., eds. Elsevier, Amsterdam
Uchida, E., Ohsumi, Y., Anraku, Y. (1985) Purification and properties of H−+-translocating, Mg−2+-adenosine triphosphatase from vacuolar membranes of Saccharomyces cerevisiae. J. Biol. Chem. 260, 1090–1095
vom Dorp, B., Scherer, G.F.E., Canut, H., Brightman, A.O., Morré, D.J. (1990) Identification of tonoplast fractions resolved from plasma membrane by free-flow electrophoresis using filipin-labeling and antibody to the tonoplast ATPase. Protoplasma 156, 57–66
Xie, X.-S., Stone, D.K. (1986) Isolation and reconstitution of the clathrin-coated vesicle proton translocating complex. J. Biol. Chem. 261, 2492–2495
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The technical assistance of Keri Safranski and Dorothy Werderitsh is gratefully acknowledged. Electron-microscope facilities were provided through the Purdue University Electron Microscope Laboratory under the direction of Professor Charles Bracker. One of the authors (G.F.E.S.) was supported by the Bundesministerium für Forschung und Technologie.
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Morré, D.J., Liedtke, C., Brightman, A.O. et al. Head and stalk structures of soybean vacuolar membranes. Planta 184, 343–349 (1991). https://doi.org/10.1007/BF00195335
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DOI: https://doi.org/10.1007/BF00195335