Summary
We recently reported the identification of a peptide (YIYGSFK) as an efficient substrate for p60c-src using a random combinatorial peptide library screening method. Over 70 analogues of YIYGSFK were designed and synthesized on beads and their phosphorylation on solid phase by p60c-src was quantitated by the PhosphorImager. A hydrophobic l-amino acid in position 2 and a basic amino acid in position 7 proved crucial for activity as a substrate. In addition, the l-tyrosine residue at position 3 was critical as the phosphorylation site and was found to be stereospecific, as substitution with the d-enantiomer at this position rendered the peptide totally inactive.
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Abbreviations
- β:
-
β-alanine
- ɛ:
-
ɛ-aminocaproic acid
- Ac:
-
N-acetyl
- BOP:
-
benzotriazol-1-yl-oxy-tris-(dimethylamino)-phosphonium hexafluorophosphate
- BSA:
-
bovine serum albumin
- Cha:
-
l-cyclohexylalanine
- Chg:
-
l-cyclohexylglycine
- Dab:
-
l-diaminobutyric acid
- Dap:
-
l-diaminoproprionic acid
- DIEA:
-
N,N-diisopropylethylamine
- DMF:
-
dimethylformamide
- Fmoc:
-
fluorenylmethyloxycarbonyl
- HOBt:
-
N-hydroxybenzotriazole
- MeF:
-
N-methyl-l-phenylalanine
- MeG:
-
N-methylglycine
- MeI:
-
N-methyl-l-isoleucine
- MES:
-
2-[N-morpholinolethanesulfonic acid
- Nle:
-
l-norleucine
- Orn:
-
l-ornithine
- TFA:
-
trifluoroacetic acid
- Z-Sar:
-
benzyloxycarbonyl-sarcosine
- Z-Tyr:
-
benzyloxycarbonyl-l-tyrosine
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Lou, Q., Wu, J., Salmon, S.E. et al. Structure-activity relationship of a novel peptide substrate for p60c-src protein tyrosine kinase. Lett Pept Sci 2, 289–296 (1996). https://doi.org/10.1007/BF00142241
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DOI: https://doi.org/10.1007/BF00142241