Abstract
A practical high-throughput protein detection system is described, based on synthetic peptide arrays consisting of designed α-helical peptides, detected by fluorescence resonance energy transfer (FRET). Initially a model α-helical peptide known to interact with a structured protein, calmodulin, was selected to establish the strategy for high-throughput detection. In comparison to peptides with a single probe, a much higher FRET response has been observed with two fluorescent probes (7-diethylaminocoumarin-3-carboxylic acid and 5(6)-carboxy-fluorescein) at both termini of the synthetic peptides. To establish a reproducible high-throughput detection system, peptides were also immobilized onto a solid surface for detection of the target proteins. A small library of 112 different peptides was constructed, based on a model of the α-helical peptide with systematic replacement of residues carrying specific charges and/or hydrophobicities. The library was used to effectively characterize various proteins, giving their own `protein fingerprint' patterns. The resulting `protein fingerprints' correlate with the recognition properties of the proteins. The present microarray with designed synthetic peptides as the capturing agents is promising for the development of protein detection chips.
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Usui, K., Takahashi, M., Nokihara, K. et al. Peptide arrays with designed α-helical structures for characterization of proteins from FRET fingerprint patterns. Mol Divers 8, 209–218 (2004). https://doi.org/10.1023/B:MODI.0000036237.82584.2d
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DOI: https://doi.org/10.1023/B:MODI.0000036237.82584.2d