2-D Phosphopeptide Mapping

  • Hikaru Nagahara
  • Robert R. Latek
  • Sergei A. Ezhevsky
  • Steven F. Dowdy
Part of the Springer Protocols Handbooks book series (SPH)


A major mechanism that cells use to regulate protein function is by phosphorylation and/or dephosphorylation of serine, threonine, and tyrosine residues. Phosphopeptide mapping of these phosphorylated residues allows investigation into the positive and negative regulatory roles these sites may play in vivo. In addition, phosphopeptide mapping can uncover the specific phosphorylated residue and, hence, kinase recognition sites, thus helping in the identification of the relevant kinase(s) and/or phosphatase(s).


Performic Acid Cooling Plate Regulate Protein Function MWCO Dialysis Tubing Supernatant Waste 
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.


  1. 1.
    Kamp, M. P. and Sefton, B. M. (1989) Acid and base hydrolysis of phosphoproteins bound to immmobilon facilitates analysis of phosphoamino acid in gel-fractionated proteins. Analyt. Biochem. 176, 22–27.CrossRefGoogle Scholar
  2. 2.
    Lees, J. A., Buchkovich, K. J., Marshak, D. R., Anderson, C. W., and Harlow, E. (1991) The retinoblastoma protein is phosphorylated on multiple sites by human cdc2. EMBO 13, 4279–4290.Google Scholar
  3. 3.
    Luo, K., Hurley, T. R, and Sefton, B. M. (1991) Cynogen bromide cleage and proteolytic peptide mapping of proteins immobilized to membranes. Meth. Enzymol. 201, 149–152.PubMedCrossRefGoogle Scholar
  4. 4.
    Desai, D., Gu, Y., and Morgan, D. O. (1992) Activation of human cyclin-dependent kinase in vitro. Mol. Biol. Cell. 3, 571–582.PubMedGoogle Scholar
  5. 5.
    Hardie, D. G. (1993) Protein Phosphorylation: A Practical Approach. IRL, Oxford, UK.Google Scholar
  6. 6.
    van der Geer, P., Luo, K., Sefton, B. M., and Hunter, T. (1993) Phosphopeptide mapping and phosphoamino acid analysis on cellulose thin-layer plates, in Protein phosphorylation-A Practical Approach. Oxford University Press, New York.Google Scholar
  7. 7.
    Harlow, E. and Lane, D. (1988) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  8. 8.
    Judd, R. C. (1994) Electrophoresis of peptides. Meth. Mol. Biol. 32, 49–57.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Hikaru Nagahara
    • 1
  • Robert R. Latek
    • 1
  • Sergei A. Ezhevsky
    • 1
  • Steven F. Dowdy
    • 1
  1. 1.Howard Hughes Medical InstituteWashington University School of MedicineSt. Louis

Personalised recommendations