Advertisement

Planta

, Volume 178, Issue 2, pp 176–183 | Cite as

Immunocytochemical localization of patatin, the major glycoprotein in potato (Solanum tuberosum L.) tubers

  • Uwe Sonnewald
  • Daniel Studer
  • Mario Rocha-Sosa
  • Lothar Willmitzer
Article

Abstract

Patatin is a family of glycoproteins with an apparent molecular weight of 40 kDa. The protein is synthesized as a pre-protein with a hydrophobic signal sequence of 23 amino acids. Using different immunocytochemical methods we determined the tissue-specific as well as subcellular localization of the patatin protein. Since antibodies raised against patatin showed crossreactivity with glycans of other glycoproteins, antibodies specific for the protein portion of the glycoprotein were purified. Using these antibodies for electron-microscopical immunocytochemistry, the protein was found to be localized mainly in the vacuoles of both tubers and leaves of potatoes (Solanum tuberosum L.) induced for patatin expression. Neither cell walls nor the intercellular space contained detectable levels of patatin protein. Concerning the tissue specificity, patatin was mainly found in parenchyma cells of potato tubers. The same distribution was observed for the esterase activity in potato tubers.

Key words

Glycoprotein Lipid acyl hydrolyse Patatin (immunocytochemical localization) Solanum (patatin localization) Vacuole 

Abbreviations

PHA

phytohemagglutinin

TFMS

trifluoromethanesulfonic acid

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ashford, D., Dwek, R.A., Welply, J.K., Hormans, S.W., Lis, H., Taylor, G.N., Radeemacher, T.W. (1987) β1→2-D-xylose and α1→3-L-fucose substituted N-linked oligosaccharides from Erythrina cristagalli lectin. Eur. J. Biochem. 166, 311–320Google Scholar
  2. Bendayan, M., Zollinger, M. (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J. Histochem. Cytochem. 31, 101–109Google Scholar
  3. Blobel, G. (1980) Intracellular protein topogenesis. Proc. Natl. Acad. Sci. USA 77, 1496–1500Google Scholar
  4. Bostock, R.M., Kuc, J., Laine, R.A. (1981) Eicosapentaenoic and arachidonic acids from Phytophtora infestans elicit fungitoxic sesquiterpenes in the potato. Science 212, 67–69Google Scholar
  5. Craig, S., Goodchild, D.J. (1984) Periodate-acid traetment of sections permits on-grid immunogold localization of pea seed vicilin in ER and Golgi. Protoplasma 122, 35–44Google Scholar
  6. Dennis, S., Galliard, T. (1974) Wax ester formation catalysed by isoenzymes of lipolytic acyl hydrolase. Phytochemistry 13, 2469–2473Google Scholar
  7. Doman, D.C., Trelease, R.N. (1985) Protein A-gold immunocytochemistry of isocitrate lyase in cotton seeds. Protoplasma 124, 157–167Google Scholar
  8. Edge, A.S.B., Faltynek, C.R., Hof, L., Reichert, L.E., Jr., Weber, P. (1981) Deglycosylation of glycoproteins by trifluoromethanesulfonic acid. Anal. Biochem. 118, 131–137Google Scholar
  9. Fournet, B., Leroy, Y., Wieruszeski, J.-M., Montreuil, J., Poretz, R.D., Goldberg, R. (1987) Prinary structure of an N-glycosidic carbohydrate unit derived from Sophora japonica lectin. Eur. J. Biochem. 166, 321–324Google Scholar
  10. Greenwood, J.S., Chrispeels, M.J. (1985) Correct targeting of the bean storage protein phaseolin in the seeds of transformed tobacco. Plant Physiol. 79, 65–71Google Scholar
  11. Kirschner, B., Hahn, H. (1986) Patatin, a major soluble protein of the potato (Solanum tuberosum L.) tuber is synthesized as a larger precursor. Planta 168, 386–389Google Scholar
  12. Laemmli, U.K. (1970) Cleavage of strcutural proteins during assembly of the head of bacteriophage T4. Nature 227, 680–685Google Scholar
  13. Mignery, G.A., Pikaard, C.S., Park, W.D. (1988) Molecular characterization of patatin multigene family of potato. Gene 62, 27–44Google Scholar
  14. Müller, M., Martin, T., Kriz, S. (1980) Improved structural preservation by: freeze-substitution. In: Electron microscopy 1980, vol 2, pp 720–721, Brederoo, T., De Priester, W., eds. 7th european congress on electron microscopy foundation, LeidenGoogle Scholar
  15. Müller, M., Moor, H. (1983) Cryofixation of thick specimes by high pressure freezing. In: The science of Biological specimen preparation for microscopy and microanalysis. Proc. 2nd Pfefferkorn Conference, pp. 131–138. Revel, J.-P., Haggis, G.H., Barnard, T. eds. SEM Inc., AMF O'HareGoogle Scholar
  16. Nishimura, M. (1982) pH in vacuoles isolated from castor bean endosperm. Plant Physiol. 70, 742–744Google Scholar
  17. Paiva, E., Lister, R.M., Park, W.D. (1983) Induction and accumulation of major tuber proteins of potato in stems and petioles. Plant Physiol. 71, 161–168Google Scholar
  18. Park, W.D., Blackwood, C., Mignery, G.A., Hermodson, M.A., Lister, R.M. (1983) Analysis of the heterogeneity of the 40,000 molecular weight tuber glycoprotein of potatoes by immunological methods and by NH2-terminal sequence analysis. Plant Physiol. 71, 156–160Google Scholar
  19. Racusen, D. (1984) Lipid acyl hydrolase of patatin. Can. J. Bot. 62, 1640–1644Google Scholar
  20. Racusen, D. (1986) Esterase specificity of patatin from two potato cultivars. Can. J. Bot. 64, 2104–2106Google Scholar
  21. Racusen, D., Foote, M. (1980) A major soluble glycoprotein of potato tubers. J. Food Biochem. 4, 43–52Google Scholar
  22. Rocha-Sosa, M., Sonnewald, U., Frommer, W., Stratmann, M., Schell, J., Willmitzer, L. (1989) Both developmental and metabolic signals activate the promoter of a class I patatin gene. EMBO J. 8, 23–29Google Scholar
  23. Rosahl, S., Eckes, P., Schell, J., Willmitzer, L. (1986) Organspecific gene expression in potato: isolation and characterization of tuber-specific cDNA sequences. Mol. Gen. Genet. 202, 368–373Google Scholar
  24. Rosahl, S., Schell, J., Willmitzer, L. (1987) Expression of a tuber specific storage protein in transgenic tobacco plants: demonstration of an esterase activity. EMBO J. 6, 1155–1159Google Scholar
  25. Smith, J.A., Hurrell, J.G.R., Leach, S.J. (1978) Elemination of nonspecific adsorption of serum proteins by sepharosebound antigens. Anal. Biochem. 87, 299–305Google Scholar
  26. Studer, D., Michel, M., Müller, M. (1989) Ciyofixation of plant tissue by high pressure freezing in: Proceedings of 9th European Congress om Electron Microscopy, York, England. in pressGoogle Scholar
  27. Sturm, A., Van Kuik, J.A., Vliegenthart, J.F.G., Chrispeels, M.J. (1987) Structure, position, and biosynthesis of the high mannose and the complex oligosaccharide side chains of the bean storage protein phaseolin. J. Biol. Chem. 262, 13392–13403Google Scholar
  28. Takahashi, N., Hotta, T., Ishihara, H., Mori, M., Tejima, S., Bligny, R., Akazawa, T., Endo, S., Arata, Y. (1986) Xylosecontaining common structural unit in N-linked oligosaccharides of laccase from sycamore cells. Biochemistry 25, 388–395Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Uwe Sonnewald
    • 1
  • Daniel Studer
    • 2
  • Mario Rocha-Sosa
    • 1
  • Lothar Willmitzer
    • 1
  1. 1.IGF BerlinBerlin 33
  2. 2.Eidgenössische Technische HochschuleMikrobiologisches InstitutZürichSwitzerland

Personalised recommendations