Abstract
Protein kinases (PKs) are a family of enzymes that catalyze the transfer of γ-phosphate from adenosine triphosphate (ATP) to tyrosine, serine, or threonine amino acid residues of substrate proteins. Phosphorylation alters the enzymatic activity, binding capability, or cellular localization of the substrate protein, as a means to relay environmental signals, such as the extracellular matrix, antigens, insulin, and growth factors (1). Following the discovery of protein phosphorylation as a mechanism of signal transduction, the discovery of the v-src and v-abl oncogenes (2,3), and the realization that PKs are an immense superfamily of proteins (2.1% of Caenor elegans genes are PKs); PKs have moved to center stage in the field of signal transduction. Because of their centrality in cell signaling, PKs have also become attractive therapeutic targets for such diverse diseases as diabetes and cancer (4).
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References
Hunter, T. (2000) Signaling-2000 and beyond. Cell 100, 113–127.
Hunter, T. and Cooper, J. A. (1985) Protein tyrosine kinases. Ann. Rev. Biochem. 54, 897–930.
Bishop, J. (1985) Viral oncogenes. Cell 42, 23–28.
Schlessinger, J. (1988) Signal transduction by allosteric receptor oligomerization. Trends Biochem. Sci. 13, 443–447.
Cooper, J. A., Esch, F. S., Taylor, S. S., and Hunter, T. (1984) Phosphorylation sites in enolase and lactate dehydrogenase utilized by tyrosine protein kinases in vivo and in vitro. J. Biol. Chem. 259, 7835–7841.
Braunwalder, A. F., Yarwood, D. R., Sills, M. A., and Lipson, K. E. (1996) Measurement of the protein tyrosine kinase activity of c-Src using time resolved fluorometry of europium chelates. Analyt. Biochem. 238, 159–164.
Braunwalder, A. F., Yarwood, D. R., Hall, T., Missbach, M., Lipson, K. E., and Sills, M. A. (1996) A solid phase assay for the determination of protein tyrosine kinase activity of c-Src using scintillating microtitration plates. Analyt. Biochem. 234, 23–26.
Wu, J., Ma, Q. N., and Lam, K. S. (1994) Identifying substrate motifs of protein kinases by a random library approach. Biochemistry 33, 14,825–14,833.
Till, J. H., Annan, R. S., Carr, S. A., and Miller, W. T. (1994) Use of synthetic peptide libraries and phosphopeptide selective mass spectrometry to probe protein kinase substrate specificity. J. Biol. Chem. 269, 7423–7428.
Zhou, S., Carraway, K. L., III, Eck, M. J., Harrison, S. C., Feldman, R. A., Mohammadi, M., et al. (1995) Catalytic specificity of protein tyrosine kinases is critical for selective signaling. Nature 373, 536–539.
Zhou, S., Blechner, S., Hoagland, N., Hoekstra, M. F., Piwnica-Worms, H., and Cantley, L. C. (1994) Use of an oriented peptide library to determine the optimal substrates of protein kinases. Curr. Biol. 4, 973–982.
Dente, L., Vetriani, C., Pelicci, G., Lanfrancone, L., Pelicci, P. G., and Cesareni, G. (1997) Modified phage peptide libraries as a tool to study specificity of phos-phorylation and recognition of tyrosine containing peptides. J. Mol. Biol. 269, 694–703.
Nishi, T., Budde, R. J., McMurray, J. S., Obeyesekere, N. U., Safdar, N., Levin, V. A., and Saya, H. (1996) Tight-binding inhibitory sequences against pp60 (c-Src) identified using a random 15 amino acid peptide library. FEBS Lett. 399, 237–240.
Schmitz, R., Baumann, G., and Gram, H. (1996) Catalytic specificity of phosphotyrosine kinases Blk, Lyn, c-Src, and Syk as assessed by phage display. J. Mol. Biol. 260, 664–677.
Chan, P. M., Keller, P. R., Connors, R. W., and Leopold, W. R., and Miller, W. T. (1996) Amino terminal sequence determinants for substrate recognition by platelet derived growth factor receptor tyrosine kinase. FEBS Lett. 394, 121–125.
Cormack, B. P., Valdivia, R. H., and Falkow, S. (1996) FACS optimized mutants of the green fluorescent protein (GFP). Gene 173, 33–38.
Shah, K., Liu, Y., Deirmengian, C., and Shokat, K. M. (1997) Engineering unnatural nucleotide specificity for Rous Sarcoma virus tyrosine kinase to uniquely label its direct substrates. Proc. Natl. Acad. Sci. USA 94, 3565–3570.
Liu, Y., Shah, K., Yang, F., Witucki, L., and Shokat, K. M. (1998) Engineering Src family protein kinases with unnatural nucleotide specificity. Chem.Biol. 5, 91–101.
Friedman, J. D. (1997) Undergraduate Thesis, Department of Molecular Biology, Princeton University, Princeton, NJ.
Dopf, J. and Horiagon, T. M. (1996) Deletion mapping of the Aequorea victoria green fluorescent protein. Gene 173, 39–44.
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Ulrich, S., Shokat, K. (2002). 22 Green Fluorescent Protein-Based Protein Kinase Biosensor Substrates. In: Hicks, B.W. (eds) Green Fluorescent Protein. Methods in Molecular Biology, vol 183. Humana Press. https://doi.org/10.1385/1-59259-280-5:275
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DOI: https://doi.org/10.1385/1-59259-280-5:275
Publisher Name: Humana Press
Print ISBN: 978-0-89603-905-6
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