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Peptide Synthesis and Applications pp 91–103Cite as

Incorporation of Phosphotyrosyl Mimetic 4- (Phosphonodifluoromethyl)phenylalanine (F2Pmp) Into Signal Transduction-Directed Peptides

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 298))

Abstract

Phosphotyrosyl (pTyr)-containing sequences in proteins serve important roles in cellular signal transduction. Often, synthetic pTyr-containing peptides based on cognate sequences surrounding these pTyr residues can exhibit pharmacologically useful properties of full phosphoproteins. However, such pTyr-containing peptides have limited use in whole-cell systems resulting from lability of the phosphate ester bond to protein-tyrosine phosphatases (PTPs). For this reason, a number of phosphatase-stable pTyr mimetics have been developed that retain certain of the chemical and pharmacological properties of pTyr itself. Among these, difluoro-phosphonomethyl phenylalanine (F2Pmp) has shown widespread utility in a variety of signal transduction settings, particularly those involving PTPs. This chapterprovides practical techniques for the synthesis of a range of F2Pmp-containing peptides.

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References

  1. Edwards, D. R. (1994) Cell signalling and the control of gene transcription. Trends Pharmacol. Sci. 15, 239–244.

    Article  PubMed  CAS  Google Scholar 

  2. Hunter, T. (2000) Signaling-2000 and beyond. Cell 100, 113–127.

    Article  PubMed  CAS  Google Scholar 

  3. Bradshaw, J. M. and Waksman, G. (2003) Molecular recognition by SH2 domains. Advances in Protein Chemistry 61, 161–210.

    Article  CAS  Google Scholar 

  4. Borg, J. P. and Margolis, B. (1998) Function of PTB domains. Protein Modules in Signal Transduction 228, 23–38.

    Article  CAS  Google Scholar 

  5. Denu, J. M. and Dixon, J. E. (1998) Protein tyrosine phosphatases: mechanisms of catalysis and regulation. Curr. Opin. Chem. Biol. 2, 633–641.

    Article  PubMed  CAS  Google Scholar 

  6. Bridges, A. (1995) The emerging role of protein phosphorylation and cell cycle control in tumor progression. Chemtracts Organic Chemistry 8, 73–107.

    CAS  Google Scholar 

  7. Cohen, P. (1999) The development and therapeutic potential of protein kinase inhibitors. Curr. Opin. Chem. Biol. 3, 459–465.

    Article  PubMed  CAS  Google Scholar 

  8. Burke, T. R. Jr., Yao, Z. J., Smyth, M. S., and Ye, B. (1997) Phosphotyrosyl-based motifs in the structure-based design of protein-tyrosine kinase-dependent signal transduction inhibitors. Curr. Pharm. Des. 3, 291–304.

    CAS  Google Scholar 

  9. Burke, T. R. Jr., Gao, Y., and Yao, Z.-J. (2000) Phosphoryltyrosyl mimetics as signaling modulators and potential antitumor agents. In Biomedical Chemistry: Applying Chemical Principles to the Understanding and Treatment of Disease, 1st ed. (Torrence, P. F., ed.), John Wiley & Sons, New York, pp. 189–210.

    Google Scholar 

  10. Burke, T. R. Jr., Yao, Z.-J., Liu, D.-G., Voigt, J., and Gao, Y. (2001) Phosphoryl-tyrosyl mimetics in the design of peptide-based signal transduction inhibitors. Biopolymers 60, 32–44.

    Article  PubMed  CAS  Google Scholar 

  11. Marseigne, I. and Roques, B. P. (1988) Synthesis of new amino acids mimicking sulfated and phosphorylated tyrosine residues. J. Org. Chem. 53, 3621–3624.

    Article  CAS  Google Scholar 

  12. Domchek, S. M., Auger, K. R., Chatterjee, S., Burke, T. R., and Shoelson, S. E. (1992) Inhibition of SH2 domain/phosphoprotein association by a nonhydrolyzable phosphonopeptide. Biochem. 31, 9865–9870.

    Article  CAS  Google Scholar 

  13. Smyth, M. S., Ford, H. Jr., and Burke, T. R. Jr. (1992) A general method for the preparation of benzylic alpha, alpha-difluorophosphonic acids; non-hydrolyzable mimetics of phosphotyrosine. Tetrahedron Lett. 33, 4137–4140.

    Article  CAS  Google Scholar 

  14. Burke, T. R. Jr., Smyth, M. S., Otaka, A., et al. (1994) Nonhydrolyzable phosphotyrosyl mimetics for the preparation of phosphatase-resistant SH2 domain inhibitors. Biochemistry 33, 6490–6494.

    Article  PubMed  CAS  Google Scholar 

  15. Burke, T. R., Kole, H. K., and Roller, P. P. (1994) Potent inhibition of insulin receptor dephosphorylation by a hexamer peptide containing the phosphotyrosyl mimetic F(2)Pmp. Biochem. Biophys. Res. Commun. 204, 129–134.

    Article  PubMed  CAS  Google Scholar 

  16. Burke, T. R. and Zhang, Z. Y. (1998) Protein-tyrosine phosphatases: Structure, mechanism, and inhibitor discovery. Biopolymers 47, 225–241.

    Article  PubMed  CAS  Google Scholar 

  17. Burke, T. R., Smyth, M. S., Nomizu, M., Otaka, A., and Roller, P. P. (1993) Preparation of fluoro-4-(phosphonomethyl)-D,L-phenylalanine and hydroxy-4-(phosphonomethyl)-D,L-phenylalanine suitably protected for solid-phase synthesis of peptides containing hydrolytically stable analogs of O-phosphotyrosine. J. Org. Chem. 58, 1336–1340.

    Article  CAS  Google Scholar 

  18. Wrobel, J. and Dietrich, A. (1993) Preparation of L-(phosphonodifluoromethyl) phenylalanine derivatives as non-hydrolyzable mimetics of O-phosphotyrosine. Tetrahedron Lett. 34, 3543–3546.

    Article  CAS  Google Scholar 

  19. Smyth, M. S. and Burke, T. R. Jr. (1994) Enantioselective synthesis of N-Boc and N-Fmoc protected diethyl 4-phosphono(difluoromethyl)-L-phenylalanine; agents suitable of the solid-phase synthesis of peptides containing nonhydrolyzable analogs of O-phosphotyrosine. Tetrahedron Lett. 35, 551–554.

    Article  CAS  Google Scholar 

  20. Qabar, M. N., Urban, J., and Kahn, M. (1997) A facile solution and solid phase synthesis of phosphotyrosine mimetic L-4-[diethylphosphono(difluoromethyl)]phenylalanine (F2Pmp(EtO)2] derivatives. Tetrahedron 53, 11,171–11,17

    Google Scholar 

  21. Otaka, A., Burke, T. R. Jr., Smyth, M. S., Nomizu, M., and Roller, P. P. (1993) Deprotection and cleavage methods for protected peptide resins containing 4-[(diethylphosphono)difluoromethyl]-DL-phenylalanine residues. Tetrahedron Lett. 34, 7039–7042.

    Article  CAS  Google Scholar 

  22. Gordeev, M. F., Patel, D. V., Barker, P. L., and Gordon, E. M. (1994) N-alpha-Fmoc-4-phosphono(difluoromethyl)-L-phenylalanine: A new O-phosphotyrosine isosteric building block suitable for direct incorporation into peptides. Tetrahedron Lett. 35, 7585–7588.

    Article  CAS  Google Scholar 

  23. Ottinger, E. A., Shekels, L. L., Bernlohr, D. A., and Barany, G. (1993) Synthesis of phosphotyrosine-containing peptides and their use as substrates for protein tyrosine phosphatases. Biochem. 32, 4354–4361.

    Article  CAS  Google Scholar 

  24. Rink, H. (1987) Solid-phase synthesis of protected peptide fragments using a trialkoxy-diphenyl-methylester resin. Tetrahedron Lett. 28, 3787–3790.

    Article  CAS  Google Scholar 

  25. Kaiser, E., Colescott, R. C., Bossinger, C. D., and Cook, P. I. (1970) Color test for the detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal. Biochem. 34, 595–598.

    Article  PubMed  CAS  Google Scholar 

  26. Pennington, M. W. (1994) Site-specific chemical modification. In Peptide Synthesis Protocols (Pennington, M. W. and Dunn, B. M., eds.), Humana Press: Totowa, NJ, p. 173.

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Medicinal Chemistry, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD

    Zhu-Jun Yao, Kyeong Lee & Terrence R. Burke Jr.

Authors
  1. Zhu-Jun Yao
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  2. Kyeong Lee
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  3. Terrence R. Burke Jr.
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Editor information

Editors and Affiliations

  1. Research Institute in Healthcare Science, School of Applied Sciences, University of Wolverhampton, Wolverhampton, UK

    John Howl

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© 2005 Humana Press Inc., Totowa, NJ

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Yao, ZJ., Lee, K., Burke, T.R. (2005). Incorporation of Phosphotyrosyl Mimetic 4- (Phosphonodifluoromethyl)phenylalanine (F2Pmp) Into Signal Transduction-Directed Peptides. In: Howl, J. (eds) Peptide Synthesis and Applications. Methods in Molecular Biology™, vol 298. Humana Press. https://doi.org/10.1385/1-59259-877-3:091

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  • DOI: https://doi.org/10.1385/1-59259-877-3:091

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-317-6

  • Online ISBN: 978-1-59259-877-9

  • eBook Packages: Springer Protocols

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