Abstract
Protein tyrosine kinases (PTKs) play prominent roles in the regulation of fundamental biological processes including normal cell growth and survival, cell differentiation, and development. Across vertebrate and invertebrate species, both nonreceptor (cytoplasmic) and receptor (transmembrane) type PTKs have been identified, making them one of the most extensively examined family of proteins. Currently, genes encoding at least 50 receptor and 33 nonreceptor vertebrate PTKs have been cloned (1,2), several by techniques that exploit the structural and functional conservation of the kinase catalytic domain.
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References
Van der Geer P. and Hunter T. (1994) Receptor protein-tyrosine kinases and their signal transduction pathways. Ann. Rev. Cell Biol. 10, 251–337.
Neet K. and Hunter T. (1996) Vertebrate non-receptor protein-tyrosine kinase families. Genes to Cells 1, 147–169.
Hanks S. H., Quinn A. M., and Hunter T. (1988) The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science 241, 42–52.
Wang J. Y. J., Queen C., and Baltimore D. (1982) Expression of an Abelson murine leukemia virus-encoded protein in Eschericia coli causes extensive phosphorylation of tyrosine residues. J. Biol. Chem. 257, 13181–13184.
Prywes R. J., Foulkes J. G., and Baltimore D. (1985) The minimum transformation region of v-abl is the segment encoding protein-tyrosine kinase. J. Virol. 54, 114–122.
Kamps M. P. and Sefton B. M. (1987) Identification of multiple novel polypeptide substrates of the v-src, v-yes, v-fps, v-ros, and v-erbB oncogenic tyrosine protein kinases utilizing antisera against phosphotyrosine. Oncogene 2, 305–515.
Stern D. F., Zheng P., Beidler D. R., and Zerillo C. (1991) Spk1, a new kinase from Saccaromyces cerevisiae, phosphorylates proteins on serine, threonine, and tyrosine. Mol. Cell. Biol. 11, 987–1001.
Mills G. B., Schmandt R., McGill M., Amendola A., Hill M., Jacobs K., May C., Rodricks A-M., Campbell S., Hogg D. (1992) Expression of TTK, a novel human protein kinase, is associated with cell proliferation. J. Biol. Chem. 267, 16,000–16,006.
Lindberg R. A., Fischer W. H., and Hunter T. (1993) Characterization of a human protein threonine kinase isolated by screening an expression library with antibodies to phosphotyrosine. Oncogene 8, 351–359.
Webster D. F., Melvin W. T., Burke M. D., and Carr E J. (1992) Screening of lambda gt1 1 cDNA libraries using monoclonal antibodies, in Methods in Molecular Biology, vol. 10, Humana, Totowa, NJ, pp. 451–460.
Snyder M., Elledge S., Sweetser D., Young R.A., and Davis R.W. (1987) Lambda gt1 1: gene isolation with antibody probes and other applications. Meth. Enzymol. 154, 107–128.
Wang J. Y. J. (1991) Generation and use of anti-phosphotyrosine antibodies raised against bacterially expressed abl protein. Meth. Enzymol. 201, 53–65.
Frackelton A. R., Posner M., Kannan B., and Mermelstein E (1991) Generation of monoclonal antibodies against phosphotyrosine and their use for affinity purification of phosphotyrosine-containing proteins. Meth. Enzymol. 201, 79–92.
Kamps M. P. (1991) Generation and use of anti-phosphotyrosine antibodies for immunoblotting. Meth. Enzymol. 201, 101–110.
Lindberg R. A. and Pasquale E. B. (1991) Isolation of cDNA clones that encode active protein-tyrosine kinases using antibodies against phosphotyrosine. Meth. Enzymol. 200, 557–564.
Tang X. X., Biegelm J. A., Nycum L. M., Yoshioka A., Brodeur G. M., Pleasure D. E., and Ikegaki N. (1995) CDNA cloning, molecular characterization, and chromosomal localization of NET(EPHT2), a human EPH-related receptor protein-tyrosine kinase gene preferentially expressed in brain. Genomics 29, 426–437.
Letwin K., Yee S.-P., and Pawson T. (1988) Novel protein-tyrosine kinase cDNAs related to fps/fes and eph cloned using anti-phosphotyrosine antibody. Oncogene 3, 621–627.
Pasquale E. B. (1990) A distinctive family of embryonic protein-tyrosine kinase receptors. Proc. Natl. Acad. Sci. USA 87, 5812–5816.
Kornbluth S., Paulson K. E., and Hanafusa H. (1988) Novel tyrosine kinase identified by phosphotyrosine antibody screening of cDNA libraries. Mol. Cell. Biol. 8, 5541–5544.
Pasquale E. B. and Singer S. J. (1989) Identification of a developmentally regulated protein-tyrosine kinase by using anti-phosphotyrosine antibodies to screen a cDNA expression library. Proc. Natl. Acad. Sci. USA 86, 5449–5453.
Takemoto Y., Sato M., Furuta M., and Hashimoto Y. (1997) Expression plasmid vectors with convenient subcloning sites in lambda gt11 that efficiently produce detectable tagged proteins. DNA Cell Biol. 16, 893–896.
Fukunaga R., and Hunter T. (1997) MNK1, a new MAP kinase-activated protein kinase, isolated by a novel expression screening method for identifying protein kinase substrates. EMBO J. 16, 1921–1933.
DiGiovanna M. P., Roussel R. R., and Stern D. F. (1995-1996) Production of antibodies that recognize specific tyrosine-phosphorylated peptides, in Curr. Prot. Prot. Sci., vol. 1, Wiley, New York, pp. 13.6.1–13.6.13.
Bangalore L., Tanner A. J., Laudano A. P., and Stern D. F. (1992) Antiserum raised against a synthetic phosphotyrosine-containing peptide selectively recognizes p185 neu/erB-2 and the epidermal growth factor receptor. Proc. Natl. Acad. Sci. USA 89, 11,637–11,641.
Sefton B. M. (1995-1996) Detection of phosphorylation by immunological techniques, in Curr. Prot. Prot. Sci., vol. 1, Wiley New York, pp. 13.4.1–13.4.5.
Gallagher S. (1995-1996) Immunoblot detection, in Curr. Prot. Prot. Sci., vol. 1, Wiley, New York, pp. 10.10.1–10.10.12.
Zhou R., Copeland T. D., Kromer L. F., and Schulz N. T. (1994) Isolation and characterization of Bsk, a growth factor receptor-like tyrosine kinase associated with the limbic system. J. Neurosci. Res. 37, 129–143.
Tabor S. and Struhl K. (1994-1997) Enzymatic Manipulation of DNA and RNA, in Curr. Prot. Molec. Biol., vol. 1, Wiley, New York, pp. 3.5.4–3.5.15.
Brown T. (1994-1997) Dot and Slot Blotting of DNA, in Curr. Prot. Molec. Biol. vol. 1, Wiley, New York, pp. 2.9.15–2.9.20.
Brown T. (1994-1997) Hybridization Analysis of DNABlots, in Curr. Prot. Molec. Biol., vol. 1, Wiley, New York, pp. 2.10.1–2.10.16.
Pasqaule E. B. (1991) Identification of chicken embryo kinase 5, a developmentally regulated receptor-type tyrosine kinse of the Eph family. Cell Reg. 2, 523–534.
Jones T. L., Karavanova I., Maeno M., Ong R. C., Kung H-F., and Daar I. O. (1995) Expression of an amphibian homolog of the Eph family of receptor tyrosine kinases is developmentally regulated. Oncogene 10, 1111–1117.
Sefton B. (1995-1996) Phosphoamino Acid Analysis, in Curr. Prot. Prot. Sci., vol. 1, Wiley, New York, pp. 13.3.1–13.3.8.
Maniatis T., Fritsch E. F., and Sambrook J. (1989) Bacteriophage vectors, in Molecular Cloning: A Laboratory Manual, vol. 1, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 2.3–2.125.
Boyle A. and Perry-O’Keefe H. (1994-1997) Labeling and colorimetric detection of nonisotopic probes, in Curr. Prot. Prot. Sci., vol. 1, Wiley, New York, pp. 3.18.1–3.18.9.
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Chong, L.D., Daar, I.O. (2000). Cloning Protein Tyrosine Kinases by Screening cDNA Libraries with Antiphosphotyrosine Antibodies. In: Reith, A.D. (eds) Protein Kinase Protocols. Methods in Molecular Biology™, vol 124. Humana Press. https://doi.org/10.1385/1-59259-059-4:21
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DOI: https://doi.org/10.1385/1-59259-059-4:21
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Print ISBN: 978-0-89603-700-7
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