, Volume 196, Issue 2, pp 284–287 | Cite as

Glycosyl-phosphatidylinositol-anchored proteins exist in the plasma membrane of Chlorella saccharophila (Krüger) Nadson: Plasma-membrane-bound nitrate reductase as an example

  • Christine Stöhr
  • Frank Schuler
  • Rudolf Tischner


Experiments with plasma-membrane vesicles were performed in order to identify the attachment of hydrophobic nitrate reductase at the plasma membrane of Chlorella saccharophila. The enzyme was successfully removed from the plasma membrane with phosphoinositol-specific phospholipase C, and showed cross-reactivity with a monoclonal antibody (clone aGPI-3) raised against the glycosyl-phosphatidylinositol (GPI) anchor of Trypanosoma variant surface protein. The enzyme was labelled in vivo by feeding [3H]ethanolamine to the cells and underwent an hydrophobicity shift after treatment with phosphoinositol-specific phospholipase C. The attachment of this form of nitrate reductase to the plasma membrane via a GPI anchor was demonstrated.

Key words

Chlorella Glycosyl-phosphatidylinositol anchor Nitrate reductase Plasma membrane 







phosphoinositol-specific phospholipase C


Plasma-membrane-bound nitrate reductase


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Cook, D.T., Clarkson, D.T., eds. (1992) Transport and receptor proteins of plant membranes. Plenum Press, New York, LondonGoogle Scholar
  2. Ferguson, M.A.J., Willams A.F. (1988) Cell surface anchoring of proteins via glycosyl-phosphatidylinositol structures. Annu. Rev. Biochem. 57, 285–320Google Scholar
  3. Hooper, N.M., Turner, A.J., eds. (1992) Lipid modifications of proteins. University Press, OxfordGoogle Scholar
  4. Larsson, C. (1985) Plasma membranes. In: Modern methods of plant analysis, vol. 1, pp. 85–104, Linkens, H.F., Jackson, J.F., eds. Springer, BerlinGoogle Scholar
  5. Lisanti, M.P., Rodriguez-Boulan, E., Saltiel, A.R. (1990) Emerging functional roles for the glycosyl-phosphatidyl-inositol membrane protein anchor. J. Membr. Biol. 117, 1–10Google Scholar
  6. Low, M.G. (1989) The glycosyl-phosphatidylinositol anchor of membrane proteins. Biochim. Biophys. Acta 988, 427–454Google Scholar
  7. Low, M.G., Saltiel, A.R.. (1988) Structural and functional roles of glycosyl-phosphatidylinositol in membranes. Science 239, 268–275PubMedGoogle Scholar
  8. Palmgren, G., Askerlund, P., Fredirikson, K., Widell, S., Sommarin, M., Larsson, C. (1990) Sealed inside-out and right-side out plasma membrane vesicles. Plant Physiol. 92, 871–880Google Scholar
  9. Preuß, U., Schuler, F., Peter-Katalinic, J., Gunawan, J., Egge, H. (1991) Production of monoclonal antibodies against a purified glycosyl-phosphatidylinositol anchor of the variant surface glycoprotein of Trypanosoma brucei. Arch. Biochem. Biophys. 291, 139–146Google Scholar
  10. Pryde, J.G. (1986) Triton X-114: a detergent that has come in from the cold. Trends Biochem. Sci. 11, 160–163Google Scholar
  11. Solomonson, L.P., Barber, M.J. (1990) Assimilatory nitrate reductase: Functional properties and regulation. Annu. Rev. Plant Physiol. Plant Mol. Biol. 41, 225–253Google Scholar
  12. Stöhr, C., Tischner, R. (1993) Characterization of the plasma-membrane-bound nitrate reductase in Chlorella saccharophila (Krüger) Nadson. Planta 191, 79–85Google Scholar
  13. Tischner, R. (1984) A comparison of the high-active and low-active form of nitrate reductase in synchronous Chlorella sorokiniana. Planta 160, 1–5Google Scholar
  14. Tischner, R., Hillmer, S., Robinson, D.G. (1987) Physiological properties and cytological features of protoplasts prepared from Chlorella saccharophila. Protoplasma 139, 153–159Google Scholar
  15. Tischner, R., Ward, M., Huffaker, R.C. (1989) Evidence for a plasma-membrane-bound nitrate reductase involved in nitrate uptake of Chlorella sorokiniana. Planta 178, 19–24Google Scholar
  16. Ward, M., Tischner, R., Huffaker, R.C. (1988) Inhibition of nitrate transport by anti-nitrate reductase IgG fragments and the identification of plasma membrane associated nitrate reductase in roots of barley seedlings. Plant Physiol. 88, 1141–1145Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Christine Stöhr
    • 1
  • Frank Schuler
    • 2
  • Rudolf Tischner
    • 1
  1. 1.Pflanzenphysiologisches InstitutUniversität GöttingenGöttingenGermany
  2. 2.Physiologisch-Chemisches InstitutUniversität BonnBonnGermany

Personalised recommendations