Abstract
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).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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. Anal. Biochem. 176, 22–27.
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.
Luo, K., Hurley, T. R, and Sefton, B. M. (1991) Cynogen bromide cleage and proteolytic peptide mapping of proteins immobilized to membranes. Methods Enzymol. 201, 149–152.
Desai, D., Gu, Y., and Morgan, D. O. (1992) Activation of human cyclin-dependent kinase in vitro. Mol. Biol. Cell. 3, 571–582.
Hardie, D. G. (1993) Protein Phosphorylation: A Practical Approach. IRL, Oxford, UK.
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.
Harlow, E. and Lane, D. (1988) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Judd, R. C. (1994) Electrophoresis of peptides. Methods Mol. Biol. 32, 49–57.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Nagahara, H., Latek, R.R., Ezhevsky, S.A., Dowdy, S.F. (1999). 2-D Phosphopeptide Mapping. In: Link, A.J. (eds) 2-D Proteome Analysis Protocols. Methods in Molecular Biology, vol 112. Humana Press. https://doi.org/10.1385/1-59259-584-7:271
Download citation
DOI: https://doi.org/10.1385/1-59259-584-7:271
Publisher Name: Humana Press
Print ISBN: 978-0-89603-524-9
Online ISBN: 978-1-59259-584-6
eBook Packages: Springer Protocols