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