Localization of In Vivo Phosphorylation Sites in Multiphosphorylated Proteins

Application of S-Ethylcysteine Derivatization and Mass Spectrometric Analysis
  • Esben S. Sørensen
  • Lone K. Rasmussen
  • Peter Højrup
  • Torben E. Petersen

Abstract

Phosphorylation of proteins is one of the most frequent forms of posttranslational modification in eukaryotic cells and is linked to the control of a multitude of cellular functions. Proteins involved in this type of regulation are typically only phosphorylated at a single or a few sites. Another type of phosphoproteins are those containing multiple phosphorylations. In these proteins the phosphorylations usually possess different functions than in proteins phosphorylated at single sites. In the case of multisite phosphorylated proteins (for review see Roach, 1991), the phosphorylations are often important as physical interactors with divalent metal ions, especially Cat2+.

Keywords

Phosphorylation Site Amino Acid Analysis Mass Spectrometric Analysis Bovine Milk Potential Phosphorylation Site 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Byford, M. S., 1991 Rapid and selective modification of phosphoserine residues catalysed by Bat2+ ions for their detection during peptide microsequencing, Biochem. J. 280: 261PubMedGoogle Scholar
  2. Holt, C., van Kemenade, M. J. J. M., 1989 The interaction of phosphoproteins with calcium phosphate, in: Calcified tissue, Hukins, D. W., ed., MacMillan, London, pp 175–213.Google Scholar
  3. Johnsen, L., Sørensen, E. S., Petersen, T. E., Berglund, L., 1995 Characterization of a bovine mammary gland PP3 cDNA reveals homology with mouse and rat adhesion molecule G1yCAM-1, Biochim. Biophys. Acta 1260: 116.PubMedCrossRefGoogle Scholar
  4. Kerr, J. M., Fisher, L. W., Termine, J. D., Young, M. F., 1991 The cDNA cloning and RNA distribution of bovine osteopontin, Gene 108: 237.PubMedCrossRefGoogle Scholar
  5. Meyer, H. E., Hoffmann-Posorske, E., Korte, H., Heilmeyer jr., M. G., 1986 Sequence analysis of phosphoserine-containing peptides. Modification for picomolar sensitivity, FEBS Lett. 204: 61.Google Scholar
  6. Meyer, H. E., Kuhn, C. C., Meyer, G. F., Swiderek, K., Weber, C., Hoffmann-Posorske, E., Heilmeyer jr., L.Google Scholar
  7. M. G., 1989 Localization of endogenous phosphoserine residues in the primary structure of proteins, in: Methods in Protein Sequence Analysis, Wittmann-Liebold, B., ed., Springer-Verlag, Berlin, pp 19–22.CrossRefGoogle Scholar
  8. Meyer, H. E., Hoffmann-Posorske, E., Heilmeyer jr., M.G., 1991 Determination and location of phosphoserine in proteins and peptides by conversion to S-ethylcysteine, Methods in Enzymology 201: 169–185.PubMedCrossRefGoogle Scholar
  9. Meyer, H. E., Eisermann, B., Heber, M., Hoffmann-Posorske, E., Korte, H., Weigt, C., Wegner, A., Hutton, T., Donella-Deana, A., Perich, J. W., 1993 Strategies for nonradioactive methods in the localization of phosphorylated amino acids in proteins, FASEB J. 7: 776.PubMedGoogle Scholar
  10. Roach, P. J., 1991 Multisite and hierarchal protein phosphorylation, J. Biol. Chem. 266: 14139.PubMedGoogle Scholar
  11. Shiraga, H., Min, W., VanDusen, W. J., Clayman, M. D., Miner, D., Terrell, C. H., Sherbotie, J. R., Forman, J. W., Przysiecki, C., Neilson, E. G., Hoyer, J. R., 1992 Inhibition of calcium oxalate crystal growth in vitro by uropontin: Another member of the aspartic acid-rich protein superfamily. Proc. Natl. Acad. Sci. USA 89: 426.PubMedCrossRefGoogle Scholar
  12. Sørensen E. S., Petersen T. E., 1993a Purification and characterization of three proteins isolated from the proteose peptone fraction of bovine milk, J. Dairy Res. 60: 189.PubMedCrossRefGoogle Scholar
  13. Sørensen E. S., Petersen T. E., 1993b Phosphoproteins, Protein Sci. 2 (Supp1.1): 111.Google Scholar
  14. Sørensen E. S., Petersen T. E., 1993c Phosphorylation, glycosylation and amino acid sequence of component PP3 from the proteose peptone fraction of bovine milk, J. Dairy Res. 60: 535.PubMedCrossRefGoogle Scholar
  15. Sørensen E. S., Petersen T. E., 1994 Identification of two phosphorylation motifs in bovine osteopontin, Biochem. Biophys. Res. Commun. 198: 200.CrossRefGoogle Scholar
  16. Sørensen E. S., Hejrup, P., Petersen, T. E., 1995 Posttranslational modifications of bovine osteopontin: Identification of twenty-eight phosphorylation and three 0-glycosylation sites, Protein Sci. 4, in press.Google Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Esben S. Sørensen
    • 1
  • Lone K. Rasmussen
    • 1
  • Peter Højrup
    • 2
  • Torben E. Petersen
    • 1
  1. 1.Protein Chemistry LaboratoryUniversity of AarhusAarhus CDenmark
  2. 2.Department of Molecular BiologyUniversity of OdenseOdense MDenmark

Personalised recommendations