biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 46:13-21, 2003 | DOI: 10.1023/A:1022341612852

Molecular Evidence for the Occurrence of H+-Transporting V-ATPase Subunit D and Two Different Forms of Subunit E in Leaves of the Obligate CAM Species Kalanchoë daigremontiana

R. Ratajczak1, T. Pfeifer1, M. Drobny1, M. Schnölzer2, U. Lüttge1
1 Institute of Botany, Darmstadt University of Technology, Darmstadt, Germany
2 German Cancer Research Center, Protein Analysis Facility, Heidelberg, Germany

Membrane proteins of purified tonoplast vesicles from leaves of Kalanchoë daigremontiana Hamet et Perrier were solubilized by the non-ionic detergent Triton X-114 and subsequently separated by MonoQ® anion-exchange chromatography. Special attention was given to the range of molecular masses around 30 kDa comprising the central stalk subunit peptides of the H+-transporting V-ATPase. Three polypeptides of apparent molecular masses of 32, 33 and 34 kDa were separated. Proteolytic fragments were obtained by trypsin digestion. Analysis by matrix-assisted laser desorption ionization (MALDI) mass spectrometry of tryptic fragments of the 32 and 33 kDa peptides and protein data- bank comparisons showed that they are two different forms of subunit E. N-terminal amino acid sequencing of tryptic fragments of the 34 kDa peptide showed that it is subunit D. This work provides for the first time unequivocal molecular evidence that the central stalk of the V-ATPase of the obligate CAM plant K. daigremontiana includes subunit D and different forms of subunit E.

Keywords: crassulacean acid metabolism; proton pump; vacuole; tonoplast
Subjects: CAM species, H+-transporting V-ATPase; Kalanchoë daigremontiana; proton pump, H+-transporting V-ATPase; tonoplast, H+-transporting V-ATPase; vacuole, H+-transporting V-ATPase

Prepublished online: March 1, 2003; Published: July 1, 2003  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Ratajczak, R., Pfeifer, T., Drobny, M., Schnölzer, M., & Lüttge, U. (2003). Molecular Evidence for the Occurrence of H+-Transporting V-ATPase Subunit D and Two Different Forms of Subunit E in Leaves of the Obligate CAM Species Kalanchoë daigremontiana. Biologia plantarum, 46(1), 13-21. doi: 10.1023/A:1022341612852
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Ames, B.N.: Assay of inorganic phosphate, total phosphate and phosphatases.-Methods Enzymol. 8: 115-118, 1966. Go to original source...
    2. Bevan, M., Pohl, T., Wcizenegger, T., Bancroft, I., Mewcs, H.W., Lemcke, K., Mayer, K.F.X.: EU Arabidopsis sequencing project.-Direct protein data base submission, accession number AAD45282: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein⌜st_uids=4850294&dopt=GenPept, 2000.
    3. Boekema, E., Ubbink-Kok, T., Lolkema, J.S., Brisson, A., Konings, W.N.: Visualization of a peripheral stalk in V-type ATPase: Evidence for the stator structure essential to rotational catalysis.-Proc. nat. Acad. Sci. USA 94: 14291-14293, 1997. Go to original source...
    4. Bordier, C.: Phase separation of integral membrane proteins in Triton X-114 solution.-J. biol. Chem. 256: 1604-1607, 1981. Go to original source...
    5. Bowman, E.J., Siebers, A., Altendorf, K.: Bafilomycins: A class of inhibitors of membrane ATPases from microorganisms, animal cells, and plant cells.-Proc. nat. Acad. Sci. USA 85: 7972-7976, 1988. Go to original source...
    6. Bremberger, C., Lüttge, U.: Tonoplast ATPase of Mesembryanthemum crystallinum. Partial amino acid sequence of subunits induced during the transition from C3-photosynthesis to crassulacean acid metabolism.-Compt. rend. Acad. Sci. Paris Sér. III 315: 119-125, 1992.
    7. Bremberger, C., Haschke, H.-P., Lüttge, U.: Separation and purification of the tonoplast ATPase and pyrophosphatase from plants with constitutive and inducible Crassulacean acid metabolism.-Planta 175: 465-470, 1988. Go to original source...
    8. Bult, C.I., White, O., Olsen, G.J., Zhou, L., Fleischmann, R.D., Sutton, G.G., Blake, J.A., Fitzgerald, L.M., Clayton, R.A., Gocayne, J.D., Kerlavagc, A.R., Dougherty, B.A., Tomb, J.-F., Adams, M.D., Reich, C.I., Overbeek, R., Kirkness, E.F., Weinstock, K.G., Merrick, J.M., Glodek, A., Scott, J.L., Geoghagen, N.S.M., Weidman, J.F., Fuhrmann, J.L., Nguyen, D., Utterback, T.R., Kelley, J.M., Peterson, J.D., Sadow, P.W., Hanna, M.C., Cotton, M.D., Roberts, K.M., Hurst, M.A., Kaine, B.P., Borodovsky, M., Klenk, H.-P., Fraser, C.M., Smith, H.O., Woese, C.R., Venter, J.C.: Complete genome sequence of the methanogenic archaeon, Methanococcus jannaschii.-Science 273: 1058-1073, 1996. Go to original source...
    9. Dietz, K.-J., Arbinger, B.: cDNA sequence and expression of subunit E of the vacuolar H+-ATPase in the inducible crassulacean acid metabolism plant Mesembryanthemum crystallinum:-Biochim. biophys. Acta 1281: 134-138, 1996. Go to original source...
    10. Dietz, K.-J., Hollenbach, B., Arnold J.: Nucleotide sequence of subunit E of the vacuolar proton-ATPase of Spinacia oleracea and Arabidopsis thaliana.-Plant Physiol. 111: 652, 1996.
    11. Dietz, K.-J., Rudloff, S., Agcorges, A., Eckerskorn, C., Fischer, K., Arbinger, B.: Subunit E of the vacuolar H+-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis.-Plant J. 8: 521-529, 1995. Go to original source...
    12. Dröse, S., Bindseil, K.U., Bowman, E.J., Siebers, A., Zeeck, A., Altendorf, K.: Inhibitory effects of modified Bafilomycins and Concanamycins on P-and V-type adenosine-triphosphatases.-Biochemistry 32: 3902-3906, 1993. Go to original source...
    13. Fischer-Schliebs, E., Ball, E., Berndt, E., Besemfelder-Butz, E., Binzel, M.L., Drobny, M., Mühlenhoff, D., Müller, M.L., Rakowski, K., Ratajczak, R.: Differential immunological cross-reactions with antisera against the V-ATPase of Kalanchoë daigremontiana reveal structural differences of V-ATPase subunits of different plant species.-Biol. Chem. 378: 1131-1139, 1997. Go to original source...
    14. Ford, C.M., Paltridge, N.G., Rathjen, J.P., Simpson, R.J., Moritz, R.L., Symons, R.H.: Assays for Yd2, the barley yellow dwarf virus resistance gene, based on the nucleotide sequence of a closely linked gene.-Direct protein data base submission, accession number AAD10336: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein&list_uids=4099150&dopt=GenPept, 1997.
    15. Gallagher, S.R., Leonhard, R.T.: Effect of vanadate, molybdate, and azide on membrane-associated ATPase and soluble phosphatase activities of corn roots.-Plant Physiol. 70: 1335-1340, 1982. Go to original source...
    16. Janbon, G., Rustchenko, E., Sherman, F.: Vacuolar ATP synthase subunit D from Candida albicans.-Direct protein data base submission, accession number P87220: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein⌜st_uids=2493133&dopt=GenPept, 1997.
    17. Jochem, P., Rona, J.-P., Smith, J.A.C., Lüttge, U.: Anion-sensitive ATPase activity and proton transport in isolated vacuoles of species of the CAM genus Kalanchoë.-Physiol. Plant. 62: 410-415, 1984. Go to original source...
    18. Kawamura, Y., Arakawa, K., Maeshima, M., Yoshida, S.: Tissue specificity of E subunit isoforms of plant vacuolar H+-ATPase and existence of isotype enzymes.-J. biol. Chem. 275: 6515-6522, 2000. Go to original source...
    19. Kim, W., Wilkins, T.A.: Isolation of a vacuolar H+-ATPase subunit E cDNA from developing cotton (Gossypium hirsutum) fibers.-Direct protein data base submission, accession number O23948: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein⌜st_uids=3334405&dopt=GenPept, 1997a.
    20. Kim, W., Wilkins, T.A.: Isolation of the vacuolar H+-ATPase subunit E cDNA from developing cotton (Gossypium hirsutum) fibers. Plant Physiol. 115: 314, 1997b.
    21. Kluge, C., Golldack, D., Dietz, K.J.: Subunit D of the vacuolar H+-ATPase of Arabidopsis thaliana.-Biochim. biophys. Acta 1419: 105-110, 1999. Go to original source...
    22. Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4.-Nature 227: 680-685, 1970. Go to original source...
    23. Lüttge, U., Fischer-Schliebs, E., Ratajczak, R., Kramer, D., Berndt, E., Kluge, M.: Functioning of the tonoplast in vacuolar C-storage and remobilization in crassulacean acid metabolism.-J. exp. Bot. 46: 1377-1388, 1995. Go to original source...
    24. Margolles-Clark, E., Tenney, K., Bowman, E.J., Bowman, B.J.: The structure of the vacuolar ATPase in Neurospora crassa.-Direct protein data base submission, accession number O59941: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein⌜st_uids=3929392&dopt=Gen Pept, 1998.
    25. Mariaux, J.-B., Becker, A., Kemna, I., Ratajczak, R., Fischer-Schliebs, E., Kramer, D., Lüttge, U., Marigo, G.: Visualization by freeze-fracture electron microscopy of intramembraneous particles corresponding to the tonoplast H+-pyrophosphatase and H+-ATPase of Kalanchoë daigremontiana Hamet et Perrier de la Bâthie.-Bot. Acta 107: 321-327, 1994. Go to original source...
    26. McIntosh, C.A., Oliver, D.J.: Isolation and characterization of the tricarboxylate transporter from pea mitochondria.-Plant Physiol. 100: 2030-2034, 1992. Go to original source...
    27. Merill, C.R., Dunau, M.L., Goldman, D.: A rapid sensitive silver stain for polypeptides in polyacrylamide gels.-Anal. Biochem. 110: 201-207, 1981. Go to original source...
    28. Mulligan, J.T., Dietrich, F.S., Hennessey, K.M., Sehl, P., Komp, C., Wei, Y., Taylor, P., Nakahara, K., Roberts, D., Davis, R.W.: Vacuolar ATP synthase subunit D of Caenorhabditis elegans.-Direct protein data base submission, accession number P32610: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein&list_uids=418399&dopt=GenPept, 1993.
    29. Nahm, B.H., Kim, J.K., Cheong, J.J., Kim, S.I., Hahn, T.R, Moon, E.P., Kim, W.T., Kim, W.Y., Yang, M.S., Park, R.D., Sohn, U.I., Kang, K.Y., Lee, M.C., Eun, M.Y.: Large-scale sequencing analysis of ESTs from rice immature seed.-Direct protein data base submission, accession number AA751826: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=nucleotide⌜st_uids=2798532&dopt=GenBank, 1998.
    30. Nelson, H., Mandiyan, S., Nelson, N.: A bovine cDNA and a yeast gene (VMA8) encoding the subunit D of the vacuolar (H+)-ATPase.-Proc. nat. Acad. Sci. USA 92: 497-501, 1995. Go to original source...
    31. Nelson, N.: Organellar Proton-ATPases.-Springer Verlag, Berlin 1995. Go to original source...
    32. Neuhoff, V., Arnold, N., Taube, D., Ehrhardt, W.: Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250.-Electrophoresis 9: 255-262, 1988. Go to original source...
    33. O'Neill, S.D., Bennett, A.B., Spanswick, R.M.: Characterization of a NO3-sensitive H+-ATPase from corn roots.-Plant Physiol. 84: 997-1000, 1983. Go to original source...
    34. Popov, N., Schmitt, M., Schulzek, S., Matthies, H.: Eine störungsfreie Mikromethode zur Bestimmung des Proteingehaltes in Gewebehomogenaten.-Acta biol. med. ger. 34: 1441-1446, 1975.
    35. Ratajczak, R.: The non-ionic detergent Brij 58 conserves the structure of the tonoplast H+-ATPase of Mesembryanthemum crystallinum during solubilization and partial purification.-Bot. Acta 107: 201-209, 1994. Go to original source...
    36. Ratajczak, R.: Structure, function and regulation of the plant vacuolar H+-translocating ATPase.-Biochim. biophys. Acta 1465: 17-36, 2000. Go to original source...
    37. Ratajczak, R., Richter, J., Lüttge, U.: Adaptation of the tonoplast V-type H+-ATPase of Mesembryanthemum crystallinum to salt stress, C3-CAM transition and plant age.-Plant Cell Environm. 17: 1101-1112, 1994. Go to original source...
    38. Rehm, H.: Der Experimentator: Proteinbiochemie.-G. Fischer Verlag, Stuttgart 1997.
    39. Reuveni, M., Sadka, A.: Isolation of vacuolar H+-ATPase subunit E cDNA from juice sacs of Citrus limon.-Direct protein data base submission, accession number AAD49706: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein⌜st_uids=5733660&dopt=GenPept, 1999.
    40. Sakai, D., Tong, H.-S., Minkin, C.: Osteoclast molecular phenotyping by random cDNA sequencing.-Bone 17: 111-119, 1995. Go to original source...
    41. Schägger, H., von Jagow, G.: Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.-Anal. Biochem. 166: 368-379, 1987. Go to original source...
    42. Serrano, R.: H+-ATPase from plasma membranes of Saccharomyces cerevisiae and Avena sativa roots: Purification and reconstitution.-Methods Enzymol. 157: 533-544, 1988. Go to original source...
    43. Smith, D.R., Doucette-Stamm, L.A., Deloughery, C., Lee, H.-M., Dubois, J., Aldredge, T., Bashirzadeh, R., Blakely, D., Cook, R., Gilbert, K., Harrison, D., Hoang, L., Keagle, P., Lumm, W., Pothier, B. Qiu, D., Spadafora, R., Vicare, R., Wang, Y., Wierzbowski, J., Gibson, R., Jiwani, N., Caruso, A., Bush, D., Safer, H., Patwell, D., Prabhakar, S., McDougall, S., Shimer, G., Goyal, A., Pietrovski, S., Church, G.M., Daniels, C.J., Mao, J.-I., Rice, P., Nolling, J., Reeve J.N.: Complete genome sequence of Methanobacterium thermoautotrophicum delta H: functional analysis and comparative genomics.-J. Bacteriol. 179: 7135-7155, 1997. Go to original source...
    44. Stapleton, A.E., Walbot, V.: Maize clones with putative DNA binding activity.-Direct protein data base submission, accession number AAD45282: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=protein⌜st_uids=5565981&dopt=GenPept, 1999.
    45. Steiger, S., Pfeifer, T., Ratajczak, R., Martinoia, M., Lüttge, U.: The vacuolar malate transporter of Kalanchoë daigremontiana: A 32 kDa polypeptide?-J. Plant Physiol. 151: 137-141, 1997. Go to original source...
    46. Stevens, T.H., Forgac, M.: Structure, function and regulation of the vacuolar (H+)-ATPase.-Annu, Rev. Cell Dev. Biol. 13: 779-808, 1997. Go to original source...
    47. Tavakoli, N., Eckerskorn, C., Golldack, D., Dietz, K.-J.: Subunit C of the vacuolar H+-ATPase of Hordeum vulgare.-FEBS Lett. 456: 68-72, 1999. Go to original source...
    48. Wilson, R., Ainscough, R., Anderson, K., Baynes, C., Berks, M., Bonfield, J., Burton, J., Connell, M., Copsey, T., Cooper, J., Coulson, A., Craxton, M., Dear, S., Du, Z., Durbin, R., Favello, A., Fraser, A., Fulton, L., Gardner, A., Green, P., Hawkins, T., Hillier, L., Jier, M., Johnston, L., Jones, M., Kershaw, J., Kirsten, J., Laisster, N., Latreille, P., Lightning, J., Lloyd, C., Mortimore, B., O'Callaghan, M., Parsons, J., Percy, C., Rifken, L., Roopra, A., Saunders, D., Shownkeen, R., Sims, M., Smaldon, N., Smith, A., Smith, M., Sonnhammer, E., Staden, R., Sulston, J., Thierry-Mieg, J., Thomas, K., Vaudin, M., Vaughan, K., Waterson, R., Watson, A., Weinstock, L., Wilkinson-Sproat, J., Wohldman, P.: 2.2 Mb of continuous nucleotide sequence from chromosome III of C. elegans.-Nature 368: 32-38, 1994. Go to original source...
    49. Zhigang, A., Löw, R., Rausch, T., Lüttge, U., Ratajczak, R.: The 32 kDa tonoplast polypeptide Di associated with the V-ATPase of Mesembryanthemum crystallinum L. in the CAM state: A proteolytically processed subunit B?-FEBS Lett. 389: 314-318, 1996. Go to original source...

    Return to the content