Abstract
The mitogen-activated protein (MAP) kinases are ubiquitous intracellular signaling proteins that respond to a variety of extracellular signals and regulate most cellular functions including proliferation, apoptosis, migration, differentiation, and secretion. The four major MAP kinase family members, which include the ERK1/2, JNK, p38, and ERK5 proteins, coordinate cellular responses by phosphorylating and regulating the activity of dozens of substrate proteins involved in transcription, translation, and changes in cellular architecture. Uncontrolled activation of the MAP kinases has been implicated in the initiation and progression of a variety of cancers and inflammatory disorders. As such, the ability to manipulate the activity of MAP kinase proteins with specific pharmacological inhibitors has received much attention as research tools for understanding basic mechanisms of cellular functions and for clinical tools to treat diseases. A variety of pharmacological inhibitors have been developed to selectively block MAP kinases directly or indirectly through targeting upstream regulators. This chapter will provide an overview of some of the current inhibitors that target MAP kinase signaling pathways and provide methodology on how to use selective MAP kinase inhibitors and immunoblotting techniques to monitor and quantify phosphorylation of MAP kinase substrates.
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References
Lewis, T. S., Shapiro, P. S., and Ahn, N. G. (1998) Signal tranduction through MAP kinase cascades. Adv Can Res 74, 49–139.
McCubrey, J. A., Milella, M., Tafuri, A., Martelli, A. M., Lunghi, P., Bonati, A., Cervello, M., Lee, J. T., and Steelman, L. S. (2008) Targeting the Raf/MEK/ERK pathway with small-molecule inhibitors. Curr Opin Investig Drugs 9, 614–630.
Cohen, P. (2009) Targeting protein kinases for the development of anti-inflammatory drugs. Curr Opin Cell Biol 21, 317–324.
Friday, B. B., and Adjei, A. A. (2008) Advances in targeting the Ras/Raf/MEK/Erk mitogen-activated protein kinase cascade with MEK inhibitors for cancer therapy. Clin Cancer Res 14, 342–346.
Kalyn, R. (2007) Overview of targeted therapies in oncology. J Oncol Pharm Pract 13, 199–205.
von Ahsen, O., and Bomer, U. (2005) High-throughput screening for kinase inhibitors. Chembiochem 6, 481–490.
Wang, Z., Canagarajah, B. J., Boehm, J. C., Kassisa, S., Cobb, M. H., Young, P. R., Abdel-Meguid, S., Adams, J. L., and Goldsmith, E. J. (1998) Structural basis of inhibitor selectivity in MAP kinases. Structure 6, 1117–1128.
Pargellis, C., Tong, L., Churchill, L., Cirillo, P. F., Gilmore, T., Graham, A. G., Grob, P. M., Hickey, E. R., Moss, N., Pav, S., and Regan, J. (2002) Inhibition of p38 MAP kinase by utilizing a novel allosteric binding site. Nat Struct Biol 9, 268–272.
Regan, J., Breitfelder, S., Cirillo, P., Gilmore, T., Graham, A. G., Hickey, E., Klaus, B., Madwed, J., Moriak, M., Moss, N., Pargellis, C., Pav, S., Proto, A., Swinamer, A., Tong, L., and Torcellini, C. (2002) Pyrazole urea-based inhibitors of p38 MAP kinase: from lead compound to clinical candidate. J Med Chem 45, 2994–3008.
Hancock, C. N., Macias, A., Lee, E. K., Yu, S. Y., Mackerell, A. D., Jr., and Shapiro, P. (2005) Identification of novel extracellular signal-regulated kinase docking domain inhibitors. J Med Chem 48, 4586–4595.
Dudley, D. T., Pang, L., Decker, S. J., Bridges, A. J., and Saltiel, A. R. (1995) A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci U S A 92, 7686–7689.
Favata, M. F., Horiuchi, K. Y., Manos, E. J., Daulerio, A. J., Stradley, D. A., Feeser, W. S., Van Dyk, D. E., Pitts, W. J., Earl, R. A., Hobbs, F., Copeland, R. A., Magolda, R. L., Scherle, P. A., and Trzaskos, J. M. (1998) Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J Biol Chem 273, 18623–18632.
Sebolt-Leopold, J. S., Dudley, D. T., Herrera, R., Van Becelaere, K., Wiland, A., Gowan, R. C., Tecle, H., Barrett, S. D., Bridges, A., Przybranowski, S., Leopold, W. R., and Saltiel, A. R. (1999) Blockade of the MAP kinase pathway suppresses growth of colon tumors in vivo. Nat Med 5, 810–816.
Kohno, M., and Pouyssegur, J. (2006) Targeting the ERK signaling pathway in cancer therapy. Ann Med 38, 200–211.
Cuenda, A., Rouse, J., Doza, Y. N., Meier, R., Cohen, P., Gallagher, T. F., Young, P. R., and Lee, J. C. (1995) SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Lett 364, 229–233.
Lee, J. C., Laydon, J. T., McDonnell, P. C., Gallagher, T. F., Kumar, S., Green, D., McNulty, D., Blumenthal, M. J., Heys, J. R., Landvatter, S. W., and et al. (1994) A protein kinase involved in the regulation of inflammatory cytokine biosynthesis. Nature 372, 739–746.
Davies, S. P., Reddy, H., Caivano, M., and Cohen, P. (2000) Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J 351, 95–105.
Bennett, B. L., Sasaki, D. T., Murray, B. W., O’Leary, E. C., Sakata, S. T., Xu, W., Leisten, J. C., Motiwala, A., Pierce, S., Satoh, Y., Bhagwat, S. S., Manning, A. M., and Anderson, D. W. (2001) SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc Natl Acad Sci U S A 98, 13681–13686.
Bain, J., McLauchlan, H., Elliott, M., and Cohen, P. (2003) The specificities of protein kinase inhibitors: an update. Biochem J 371, 199–204.
Tatake, R. J., O’Neill, M. M., Kennedy, C. A., Wayne, A. L., Jakes, S., Wu, D., Kugler, S. Z., Jr., Kashem, M. A., Kaplita, P., and Snow, R. J. (2008) Identification of pharmacological inhibitors of the MEK5/ERK5 pathway. Biochem Biophys Res Commun 377, 120–125.
Arora, A., and Scholar, E. M. (2005) Role of tyrosine kinase inhibitors in cancer therapy. J Pharmacol Exp Ther 315(3), 971–979.
Mendelsohn, J., and Baselga, J. (2006) Epidermal growth factor receptor targeting in cancer. Semin Oncol 33, 369–385.
Hahn, S. M., Bernhard, E., and McKenna, W. G. (2001) Farnesyltransferase inhibitors. Semin Oncol 28, 86–93.
Mesa, R. A. (2006) Tipifarnib: farnesyl transferase inhibition at a crossroads. Expert Rev Anticancer Ther 6, 313–319.
Wong, S., and Witte, O. N. (2004) The BCR-ABL story: bench to bedside and back. Annu Rev Immunol 22, 247–306.
Bocchia, M., Forconi, F., and Lauria, F. (2006) Emerging drugs in chronic myelogenous leukaemia. Expert Opin Emerg Drugs 11, 651–664.
Mody, N., Leitch, J., Armstrong, C., Dixon, J., and Cohen, P. (2001) Effects of MAP kinase cascade inhibitors on the MKK5/ERK5 pathway. FEBS Lett 502, 21–24.
Acknowledgements
The authors would like to thank Kimberly Still for technical assistance. This work was supported by the National Institutes of Health (CA120215).
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Burkhard, K., Shapiro, P. (2010). Use of Inhibitors in the Study of MAP Kinases. In: Seger, R. (eds) MAP Kinase Signaling Protocols. Methods in Molecular Biology, vol 661. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-795-2_6
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DOI: https://doi.org/10.1007/978-1-60761-795-2_6
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