Identification of GIYWHHY as a novel peptide substrate for human p60c-src protein tyrosine kinase
✍ Scribed by Qiang Lou; Margaret E. Leftwich; Kit S. Lam
- Publisher
- Elsevier Science
- Year
- 1996
- Tongue
- English
- Weight
- 600 KB
- Volume
- 4
- Category
- Article
- ISSN
- 0968-0896
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✦ Synopsis
We have recently determined that -Ile-Tyr- were the two critical residues as a peptide substrate for p60c-src protein tyrosine kinase (Lou, Q. et al., Lett. Peptide Sci., 1995, 2, 289). Here, we report on the design and synthesis of a secondary 'one-bead, one-compound' combinatorial peptide library based on this dipeptide motif (XIYXXXX, where X = all 19 eukaryotic amino acids except for cysteine). This secondary library was screened for its ability to be phosphorylated by p60c-src PTK using [gamma 32P]ATP as a tracer. Five of the strongest [32P]-labeled peptide-beads were identified and microsequenced: GIYWHHY, KIYDDYE, EIYEENG, EIYEEYE, and YIYEEED. A solid-phase phosphorylation assay was used to evaluate the structure-activity relationship of GIYWHHY. It was determined that Ile2, Tyr3, His5, and His6 were crucial for its activity as a substrate.