Use of intein-directed peptide biosynthesis to improve serum stability and bioactivity of a gelatinase inhibitory peptide
| Autor: | Heli Valtanen, Harri Savilahti, Erkki Koivunen, Mikael Björklund |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Gelatinases
Phage display Recombinant Fusion Proteins Peptide Biology 03 medical and health sciences 0302 clinical medicine Drug Stability Cell Movement Peptide Library Drug Discovery Tumor Cells Cultured Gelatinase Humans Protein Splicing Peptide Biosynthesis Amino Acid Sequence Enzyme Inhibitors Peptide library Peptide sequence 030304 developmental biology chemistry.chemical_classification 0303 health sciences Alanine Organic Chemistry Tryptophan General Medicine 3. Good health Computer Science Applications Mutagenesis Insertional Biochemistry chemistry 030220 oncology & carcinogenesis Intein Peptides |
| Zdroj: | Combinatorial chemistryhigh throughput screening. 6(1) |
| ISSN: | 1386-2073 |
| Popis: | Screening of phage display libraries allows rapid identification of peptides binding to a target. However, functional analysis of the phage sequences and their reproduction as soluble and stable peptides are often the most time-consuming part in the screening. We have used here intein-based peptide biosynthesis to produce a phage-display derived gelatinase inhibitory peptide CTTHWGFTLC and to identify the critical residues for gelatinase inhibitory activity by performing alanine-scanning mutagenesis. By biosynthetic incorporation of 5-fluorotryptophan, we obtained an inhibitor of MMP-2 and MMP-9 gelatinases that showed a 6-fold enhancement in serum stability in comparison to the wild-type peptide. The new peptide also had an improved ability to inhibit tumor cell migration. These studies indicate the utility of intein methodology for synthesis and design of peptides obtained by phage display. |
| Databáze: | OpenAIRE |
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