Fmoc-based chemical synthesis and selective binding to supercoiled DNA of the p53 C-terminal segment and its phosphorylated and acetylated derivatives
✍ Scribed by Kenta Teruya; Angela C. Murphy; Tom Burlin; Ettore Appella; Sharlyn J. Mazur
- Book ID
- 105360363
- Publisher
- John Wiley and Sons
- Year
- 2004
- Tongue
- English
- Weight
- 260 KB
- Volume
- 10
- Category
- Article
- ISSN
- 1075-2617
- DOI
- 10.1002/psc.552
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✦ Synopsis
Abstract
The C‐terminal domain of p53 comprises a linker, the tetramerization domain and the regulatory domain, and contains at least seven sites of potential post‐translational modification. An improved strategy was developed for the synthesis of large peptides that contain phosphorylated amino acids and p53(303–393), a 91‐amino acid peptide, and three post‐translationally modified derivatives were synthesized through the sequential condensation of three partially protected segments. Peptide thiolesters were prepared using the sulfonamide‐based ‘safety‐catch’ resin approach and employing Fmoc‐based solid‐phase peptide synthesis. At the N‐terminus of the middle building block, a photolabile protecting group, 3,4‐dimethoxy‐6‐nitrobenzyloxycarbonyl, was incorporated to differentiate the N‐terminal amino group from the side‐chain amino groups. Two sequential couplings were accomplished following this protection strategy. The synthetic products, p53(303–393) and its phosphorylated or acetylated derivatives, exhibited the ability to bind specifically to supercoiled DNA, which is one of the characteristics of this domain. Published in 2004 by the European Peptide Society and John Wiley & Sons, Ltd.