A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Use of intein-directed peptide biosynthesis to improve serum stability and bioactivity of a gelatinase inhibitory peptide
Tekijät: Bjorklund M, Valtanen H, Savilahti H, Koivunen E
Kustantaja: BENTHAM SCIENCE PUBL LTD
Julkaisuvuosi: 2003
Lehti: Combinatorial Chemistry and High Throughput Screening
Tietokannassa oleva lehden nimi: COMBINATORIAL CHEMISTRY & HIGH THROUGHPUT SCREENING
Lehden akronyymi: COMB CHEM HIGH T SCR
Vuosikerta: 6
Numero: 1
Aloitussivu: 29
Lopetussivu: 35
Sivujen määrä: 7
ISSN: 1386-2073
Tiivistelmä
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.
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.
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