Skip to main content
Log in

Development of a selective pseudosubstrate-based peptide inhibitor of pp60c-src protein tyrosine kinase

Letters in Peptide Science Aims and scope Submit manuscript

Summary

Using a combinatorial peptide library method, we identified YIYGSFK as an efficient and specific peptide substrate for pp60c-src protein tyrosine kinase (PTK) [Lam et al., Int. J. Pept. Protein Res., 45 (1995) 587]. Employing YIYGSFK as a template, we synthesized and evaluated a series of pseudosubstrate-based inhibitors for pp60c-src. We found that the efficiency of a given inhibitor was highly dependent on the specific tyrosine analog used at the phosphorylation site of the substrate. One of these pseudosubstrate inhibitors, YI(2′-Nal)GSFK, selectively inhibited the kinase activity of pp60c-src, with a Ki of 24 μM. This peptide inhibitor exhibited selectivity for pp60c-src as compared to other PTKs tested, such as c-Abl and Bcr-Abl. Our results suggest that selective inhibitors for a specific PTK can be developed when the structure of a specific and efficient small peptide substrate for this PTK can be used as a template for structure modification.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Abbreviations

1′-Nal:

l-1′-naphthylalanine

2′-Nal:

l-2′-naphthylalanine

BOP:

benzotriazolyl-N-oxy-tris(dimethylamino)-phosphonium hexafluorophosphate

BSA:

bovine serum albumin

cAPK:

cyclic AMP-dependent protein kinase

DIEA:

diisopropylethylamine

EGFR:

epidermal growth factor receptor

Fmoc:

fluorenylmethoxycarbonyl

HOBt:

1-hydroxybenzotriazole

MES:

2-[N-morpholino]ethanesulfonic acid

PBS:

phosphate-buffered salts

pCl:

l-p-chlorophenylalanine

pF:

l-p-fluorophenylalanine

PTK:

protein tyrosine kinase

TLC:

thin-layer chromatography

References

  1. Hunter, T., Curr. Opin. Cell Biol., 1 (1989) 1168.

    Google Scholar 

  2. Fantl, W.J., Johnson, D.E. and Williams, L.T., Annu. Rev. Biochem., 62 (1993) 453.

    Google Scholar 

  3. Chan, A.C., Desai, D.M. and Weiss, A., Annu. Rev. Immunol., 12 (1994) 555.

    Google Scholar 

  4. Mustelin, T. and Burn, P., Trends Biochem. Sci., 18 (1993) 215.

    Google Scholar 

  5. White, M.F.J., Bioenerg. Biomembrane, 23 (1991) 63.

    Google Scholar 

  6. Levitzki, A. and Gazit, A., Science, 267 (1995) 1782.

    Google Scholar 

  7. Ross, R., Nature, 362 (1993) 801.

    Google Scholar 

  8. Elder, J.T., Fisher, G.J., Lindquist, P.B., Bennett, G.L., Pittelkow, M.R., CoffeyJr., R.J., Ellingsworth, L., Derynck, R. and Voorhees, J.J., Science, 243 (1989) 811.

    Google Scholar 

  9. Cook, P.W., Pittelkow, M.R., Keeble, W.W., Graves-Deal, R., Coffey, R.J. and Shipley, G.D., Cancer Res. 52 (1992) 3224.

    Google Scholar 

  10. Burke, T.R., Drugs Future, 17 (1992) 119.

    Google Scholar 

  11. Fry, D.W., Kraker, A.J., McMichael, A., Ambroso, L.A., Nelson, J.M., Leopold, W.R., Connors, R.W. and Bridges, A.J., Science, 265 (1994) 1093.

    Google Scholar 

  12. Hagiwara, M., Inoue, S., Tanaka, T., Nunoki, K., Ito, M. and Hidaka, H., Biochem. Pharmacol., 37 (1988) 2987.

    Google Scholar 

  13. Bolen, J.B., Rosen, N. and Israel, M.A., Proc. Natl. Acad. Sci. USA, 82 (1985) 7275.

    Google Scholar 

  14. Bolen, J.B., Veillette, A., Schwartz, A.M., Deseau, V. and Rosen, N., Proc. Natl. Acad. Sci. USA, 84 (1987) 2251.

    Google Scholar 

  15. Cartwright, C.A., Kamps, M.P., Meisler, A.I., Pipas, J.M. and Eckhard, W., J. Clin. Invest., 83 (1989) 2025.

    Google Scholar 

  16. Cartwright, C.A., Meisler, A.I. and Eckhard, W., Proc. Natl. Acad. Sci. USA, 87 (1990) 558.

    Google Scholar 

  17. Rosen, N., Bolen, J.B., Schwartz, A.M., Cohen, P., Deseau, V. and Israel, M., J. Biol. Chem., 261 (1986) 13754.

    Google Scholar 

  18. Talamonti, M.S., Curley, S.A. and Gallick, G.E., Cancer Bull., 44 (1992) 321.

    Google Scholar 

  19. Talamonti, M.S., Roh, M.S., Curley, S.A. and Gallick, G.E., J. Clin. Invest., 91 (1993) 53.

    Google Scholar 

  20. Termuhlen, P.M., Curley, S.A., Talamonti, M.S., Saboorian, M.H. and Gallick, G.E., J. Surg. Res., 54 (1993) 293.

    Google Scholar 

  21. Ottenhoff-Kalff, A.E., Rijksen, G., Van Beurden, A.A.C.M., Hennipman, A., Michels, A.A. and Staal, G.E.J., Cancer Res., 52 (1992) 4773.

    Google Scholar 

  22. Partanen, S., Oncology Rep., 1 (1994) 603.

    Google Scholar 

  23. Takeshima, E., Hamaguchi, M., Watanabe, T., Akiyama, S., Ohnishi, Y., Xiao, H., Nagai, Y. and Takaagi, H., Jpn. J. Cancer Res., 82 (1991) 1428.

    Google Scholar 

  24. O'Shaughnessy, J., Deseau, V., Amini, S., Rosen, N. and Bolen, J.B., Oncogene Res., 2 (1987) 1.

    Google Scholar 

  25. Bjelfman, C., Hedborg, F., Johansson, I., Nordenskjold, M. and Pahlman, S., Cancer Res., 50 (1990) 6908.

    Google Scholar 

  26. Fanning, P., Bulovas, K., Saini, K.S., Libertino, J.A., Joyce, A.D. and Summerhayes, I.C., Cancer Res., 52 (1992) 1457.

    Google Scholar 

  27. Lynch, S.A., Brugge, J.S., Fromowitz, F., Glantz, L., Wang, P., Caruso, R. and Viola, M.V., Leukemia, 7 (1993) 1416.

    Google Scholar 

  28. Talamonti, M.S., Roh, M.S., Curley, S.A. and Gallick, G.E., Surg. Form., 42 (1991) 422.

    Google Scholar 

  29. Jessup, J.M. and Gallick, G.E., Curr. Problems Cancer, 16 (1992) 263.

    Google Scholar 

  30. Lam, K.S., Salmon, S.E., Hersh, E.M., Hruby, V.J., Kazmierski, W.M. and Knapp, R.J., Nature, 354 (1991) 82.

    Google Scholar 

  31. Wu, J., Ma, Q.N. and Lam, K.S., Biochemistry, 33 (1994) 14825.

    Google Scholar 

  32. Lam, K.S., Wu, J. and Lou, Q., Int. J. Pept. Protein Res., 45 (1995) 587.

    Google Scholar 

  33. Stewart, J.M. and Young, J.D. (Eds.) Solid Phase Peptide Synthesis, Pierce Chemical Co., Rockford, IL, U.S.A., 1984.

    Google Scholar 

  34. Atherton, E. and Sheppard, R.C. (Eds.) Solid Phase Peptide Synthesis, IRL Press at Oxford University Press, Oxford, U.K., 1989.

    Google Scholar 

  35. Lou, Q., Wu, J. and Lam, K.S., Anal. Biochem., 235 (1996) 107.

    Google Scholar 

  36. Songyang, Z., CarrawayIII, K.L., Eck, M.J., Harrison, S.C., Feldman, R.A., Mohammadi, M., Schlessinger, J., Hubbard, S.R., Smith, D.P., Eng, C., Lorenzo, M.J., Ponder, B.A.J., Mayer, B.J. and Cantley, L.C., Nature, 373 (1995) 536.

    Google Scholar 

  37. Nair, S.A., Kim, M.H., Warren, S.D., Choi, S., Songyang, Z., Cantley, L.C. and Hangauer, D.G, J. Med. Chem., 38 (1995) 4276.

    Google Scholar 

  38. Cheng, H.-C., Litwin, C.M.E., Hwang, D.M. and Wang, J.H., J. Biol. Chem., 266 (1991) 17919.

    Google Scholar 

  39. Cheng, H.-C., Nishio, H., Hatase, O., Ralph, S. and Wang, J.H., J. Biol. Chem., 267 (1992) 9248.

    Google Scholar 

  40. Cheng, H.-C., Matsuura, I. and Wang, J.H., Mol. Cell. Biochem., 127/128 (1993) 103.

    Google Scholar 

  41. Yabe, Y., Miura, C., Horikoshi, H. and Baba, Y., Chem. Pharm. Bull., 24 (1976) 3149.

    Google Scholar 

  42. Ljungqvist, A., Feng, D.M., Bowers, C., Hook, W.A. and Folkers, K., Tetrahedron, 46 (1990) 3297.

    Google Scholar 

  43. Rodriquez, M., Bernad, N., Galas, M.C., Lignon, M.F., Laur, J., Aumelas, A. and Martinez, J., Eur. J. Med. Chem., 26 (1991) 245.

    Google Scholar 

  44. Darlak, K., Miller, R.B., Stack, M.S., Spatola, A.F. and Gray, R.D., J. Biol. Chem., 265 (1990) 5199.

    Google Scholar 

  45. Yuan, C.J., Jakes, S., Elliot, S. and Graves, D.J., J. Biol. Chem., 265 (1990) 16205.

    Google Scholar 

  46. Fry, D.W., McMichael, A., Singh, J., Dobrusin, E.M. and McNamara, D.J., Peptides, 15 (1994) 951.

    Google Scholar 

  47. Hanks, S.K., Quinn, A.M. and Hunter, T., Science, 241 (1988) 42.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, J.J., Phan, H., Salmon, S.E. et al. Development of a selective pseudosubstrate-based peptide inhibitor of pp60c-src protein tyrosine kinase. Lett Pept Sci 3, 309–316 (1996). https://doi.org/10.1007/BF00127665

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00127665

Keywords

Navigation